Pharmaceutical preparations and methods to manage weight and to modulate the gut microbiota

ABSTRACT

A pharmaceutical preparation, comprising: A) A plurality of particles, each particle comprising: 1) a core comprising at least one agent that increases oxygen tension and/or redox potential and/or pH in the colon of a subject following administration to the subject, and 2) an enteric coating encasing the core; and B) a capsule or sachet encasing the plurality of particles. Methods of weight management for a subject, comprising administering the pharmaceutical preparation to a subject in need thereof.

INTRODUCTION

With only diet, exercise, and bariatric surgery as options, the lack ofeffective and safe medications for treating metabolic disorders anddiseases (such as obesity), metabolic syndrome, and cardiovasculardisease are significant unmet medical needs. For example, obesity is alife-threatening disorder in which there is an increased risk ofmorbidity and mortality arising from concomitant diseases such as typeII diabetes, hypertension, obstructive sleep apnea, inflammation,stroke, cancer and gallbladder disease. Obesity is now a majorhealthcare issue in the Western World and increasingly in some thirdworld countries. Currently about 35% of the population of the USA is nowconsidered obese and an additional ˜40% overweight. Management of bodyweight and body mass index (BMI) is also correlated with many indicatorsof health and even a numerically small reduction of body weight can leadto a profound improvement in obesity related conditions and increasequality of life and life expectancy.

Microbiome targeted therapeutics have a potential to address thisproblem, as it is becoming increasingly clear that the gut microbiotahas a causal role in alleviating or mediating metabolic disorders,including obesity. However, microbiome modulation approaches to datehave predominantly focused on utilizing probiotics, prebiotics,antibiotics or fecal transplants, which lack targeted or broad-spectrumefficacy and may have unfavorable safety and efficacy profiles.Accordingly, there is a need for new methods and compositions formanaging the weight of a subject and/or treating metabolic disordersincluding obesity, metabolic syndrome, and cardiovascular disease. Theinventions disclosed herein meet these and other needs.

SUMMARY

The inventor has discovered that administering an effective amount tothe small intestine and/or large intestine of a subject of at least oneagent that increases oxygen tension and/or redox potential and/or pH inthe colon of the subject has a beneficial effect on the gut microbiomeof the subject. By increasing the oxygen tension and/or redox potentialand/or pH the colon environment is modified such that (1) the relativeabundance of bacterial types known to promote at least one conditionselected from a metabolic disease or disorder (such as obesity),metabolic syndrome, and cardiovascular disease is reduced; and/or (2)the relative abundance of bacterial types known to ameliorate at leastone condition selected from a metabolic disease or disorder (such asobesity), metabolic syndrome, and cardiovascular disease is increased.Modulation in this way provides a means to manage the weight of asubject. See for example co-pending International Application No.PCT/US2016/034973, filed 31 May 2016.

This invention specifically provides improved pharmaceuticalpreparations for delivering the at least one agent that increases oxygentension and/or redox potential and/or pH in the colon of a subject tothe target area within the gastrointestinal tract of the subject. Theinventor has found that a multiparticulate pharmaceutical preparation inwhich each particule is enteric coated and then the particles areencased in a capsule or sachet for administration to a subject providesa desirable means of delivering the at least one agent that increasesoxygen tension and/or redox potential and/or pH in the colon of asubject to the target area within the gastrointestinal tract of thesubject. In several embodiments this formulation exhibits advantagesrelative to alternatives in which an entire dose of the agent is encasedin a enteric coating (for example as an enteric coated tablet or anenteric coated capsule (in which the enteric coating is on the outsideof the capsule).

For example capsules are hard to coat reliably with enteric coating. Ifcoating thickness is too low, then the capsules fail to reliably showenteric properties (they open in acidic conditions). This is due to afailure point at the capsule junction where the body and the cap of thecapsule connect. A thin layer of coating sometimes fails to coat overthis gap causing failure.

On the other hand, if the coating thickness is increased, it is possibleto apply a large amount of polymer over this gap to coat it morereliably. However, unfortunately, a high coating thickness also leads toa non-functional formulation, because the capsules fail to open andrelease API in vivo. A thick coating layer fails to dissolve in time toopen before reaching the colon.

In certain formulations of the invention small round spherical particlesof active agent are coated, for example using a fluidized bed. Thisapproach allows coating of particles with smooth surfaces and mitigatesany coating failure due to irregularities on the surface (such as acap-body junction of capsule). This further enables a low enough coatingthickness such that the formulation is both acid resistant and opensrapidly once the correct pH is reached in the distal small intestine.

In certain embodiments this approach also provides one or more of thefollowing advantages: Particles spread uniformly over the GI tract,which decrease local concentration of API in the intestines and create asustained delivery of API to the proximal colon over 3-5 hours; andlower local concentration of API helps reducing any GI irritation orside effects.

Accordingly, in a first aspect this invention provides pharmaceuticalpreparations comprising a plurality of particles, each particlecomprising: 1) a core comprising at least one agent that increasesoxygen tension and/or redox potential and/or pH in the colon of asubject following administration to the subject, and 2) an entericcoating encasing the core.

In some embodiments the pharmaceutical preparation is a unit dosageform. In some embodiments the unit dosage form consists of a knownnumber of particles.

In some embodiments the pharmaceutical preparations comprise a pluralityof particles and a capsule or sachet encasing the plurality ofparticles. In some embodiments the pharmaceutical preparation comprises:A) A plurality of particles, each particle comprising: 1) a corecomprising at least one agent that increases oxygen tension and/or redoxpotential and/or pH in the colon of a subject following administrationto the subject, and 2) a delayed release coating encasing the core; andB) a capsule or sachet encasing the plurality of particles.

In some embodiments the at least one agent is selected from carbamideperoxide and sodium percarbonate. In some embodiments the at least oneagent is sodium percarbonate. In some embodiments the core comprisescatalase. In some embodiments the particles further comprise a releaserate modifier. In some embodiments the core of the particles comprisesthe release rate modifier. In some embodiments the particles furthercomprise a subcoat layer that encases the core and is disposed betweenthe core and the delayed release coating. In some embodiments thesubcoat layer comprises HPMC

In some embodiments the weight ratio of delayed release coating to coreof the particles is from 5% to 50%. In some embodiments the weight ratioof delayed release coating to core of the particles is from 20% to 30%.In some embodiments the weight ratio of delayed release coating to coreof the particles is from 30% to 40%. In some embodiments the weightratio of delayed release coating to core of the particles is from 40% to50%.

In some embodiments the delayed release coating is an enteric coatingthat is protective at pH below 5.5. In some embodiments the entericcoating is protective at pH below 6.5. In some embodiments the entericcoating is protective at pH below 7.0. In some embodiments the entericis protective at pH below 7.4. In some embodiments the delayed releasecoating is Eudagrit FS30D.

In some embodiments, following oral administration of the pharmaceuticalpreparation to a subject the core is delivered to the distal ileumand/or colon.

In some embodiments the cores of the plurality of particles have amaximum dimension of from 0.1 to 2 mm. In some embodiments the cores ofthe plurality of particles have a maximum dimension of from 0.2 to 1.0mm. In some embodiments the plurality of particles have a maximumdimension of 3.0 mm or less.

In some embodiments the pharmaceutical preparation is provided in a unitdosage form comprising from 25 to 500 mg of the at least one agent. Insome embodiments the pharmaceutical preparation is provided in a unitdosage form comprising from 50 to 200 mg of the at least one agent.

This invention also provides methods of using the pharmaceuticalpreparations of the invention for weight management of a subject. Insome embodiments the methods comprise administering a pharmaceuticalpreparation of the invention to the subject. In some embodiments weightmanagement comprises at least one of weight loss, maintenance of weight,controlling weight gain, body mass index (BMI) reduction, maintenance ofBMI, and controlling BMI gain. In some embodiments the subject isoverweight or obese. In some embodiments the subject has at least oneweight-related condition. In some embodiments the at least oneweight-related condition is selected from a metabolic disease ordisorder, metabolic syndrome, and cardiovascular disease.

In some embodiments administering the pharmaceutical preparation to thesubject increases the relative abundance of at least one of aerobicbacteria and facultatively anaerobic bacteria in the microbiota of thecolon of the subject by at least 100%. In some embodiments administeringthe pharmaceutical preparation to the subject increases the relativeabundance of bacteria in at least one phylum selected fromProteobacteria, Bacteriodetes, and Verrucomicrobia in the microbiota ofthe colon of the subject by at least 100%. In some embodimentsadministering the pharmaceutical preparation to the subject decreasesthe relative abundance of bacteria in the Firmicutes phylum in themicrobiota of the colon of the subject.

In some embodiments the pharmaceutical preparation is administered oncea day, twice a day, three times a day, or four times a day. In someembodiments from 50 to 200 mg of the at least one agent is administeredat each dosing. In some embodiments the pharmaceutical preparation isadministered for a dosing period of at least five days, and wherein theweight of the subject in reduced by at least 2%. In some embodiments thepharmaceutical preparation is administered for a dosing period of atleast five days, and wherein the weight of the subject in reduced by atleast 5%.

In some embodiments the subject is a human.

In some embodiments administering the pharmaceutical preparation to thesubject increases ketone body excretion in the urine of the subject.

This invention also provides methods of increasing ketogenesis by asubject. In some embodiments the methods comprise administering apharmaceutical preparation of the invention to the subject. Thisinvention also provides uses of a pharmaceutical preparation of theinvention to manage the weight of a subject.

This invention also provides uses of a pharmaceutical preparation of theinvention for manufacturing a medicament intended to manage the weightof a subject.

This invention also provides uses of a pharmaceutical preparation of theinvention to increase ketogenesis by a subject.

This invention also provides uses of a pharmaceutical preparation of theinvention for manufacturing a medicament intended to increaseketogenesis by a subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows bodyweight time courses for subjects 1-3.

FIG. 2 shows fasting SMPG and urine ketone results for subjects 1 and 2.Each fasting SMPG data point indicates the average of plasma glucosemeasurements over the previous 8-days.

FIG. 3 shows urine ketone results for subject 3. Urine ketone levels(acetoacetate) increased in subject 3 during dosing as measured bystandard urine ketone strips.

FIG. 4 shows additional body weight data for subject 3.

FIG. 5 shows body weight timecourses for rats.

DETAILED DESCRIPTION A. Introduction

Bariatric surgical procedures such as vertical sleeve gastrectomy (VSG)and Roux-en-Y gastric bypass (RYGB) are the most potent treatmentsavailable to produce sustained reductions in body weight andimprovements in glucose regulation. While traditionally these effectsare attributed to mechanical aspects of these procedures, such asrestriction and malabsorption, a growing body of evidence from mousemodels of these procedures points to physiological changes that mediatethe potent effects of these surgeries. In particular, there are similarchanges in gut hormone secretion, bile acid levels, and compositionafter both of these procedures. Moreover, loss of function of thenuclear bile acid receptor (FXR) greatly diminishes the effects of VSG.Bariatric surgeries are linked to profound changes in the gut microbiomethat also mediate at least some of these surgical effects.

Intestinal microorganisms could be contributing to obesity by increasingrecovery of energy from the diet and via the impact of microbialmetabolites or microbial cell-derived signals on host pathways thatregulate energy homoeostasis and lipid metabolism. Several gut microbialdiversity surveys in mouse models [Ley R E, Backhed F, Turnbaugh P,Lozupone C A, Knight R D, Gordon J I. “Obesity alters gut microbialecology.” Proc Natl Acad Sci USA 2005; 102: 11070-11075] and humans [LeyR E, Turnbaugh P J, Klein S, Gordon J I. “Microbial ecology: human gutmicrobes associated with obesity.” Nature 2006; 444: 1022-1023; andTurnbaugh P J, Hamady M, Yatsunenko T, Cantarel B L, Duncan A, Ley R Eet al. “A core gut microbiome in obese and lean twins.” Nature 2009;457: 480-484] provided evidence that obesity was associated with adecreased proportion of Bacteroidetes and a higher proportion ofFirmicutes. These and other data suggest that the gut microbiome plays aregulatory role in obesity and related metabolic conditions.

To date, efforts to modulate the gut microbiome have focused onstrategies to directly add helpful types of bacteria to the gut and/orstrategies to reduce harmful types of bacteria in the gut by the use ofantibiotics. These approaches are necessarily challenging because addedbacteria must compete against endogenous bacteria and antibiotics areoften a crude tool that target several types of bacteria at once andthere are concerns over antibiotic resistance development. Accordingly,there is a need in the art to provide methods and compositions to modifythe gut microbiome in a manner that counters obesity and other metabolicconditions and favors a healthy glucose balance.

U.S. Pat. No. 5,759,539 in the name of David R. Whitmire, issued Jun. 2,1998 (“Whitmore”) suggests that oxygen generating formulations such ascatalase and hydrogen peroxide may be useful in combination with enzymesthat oxidize ethanol to acetate to treat ethanol overdose and to reducethe amount of ethanol in a subject. Whitmore suggests that oxygen isadministered directly with a carrier or that an oxygen generator isadministered. In Whitmore the oxygen or oxygen generator is administeredtogether with enzymes that oxidize ethanol to acetate, and thesecomponents are delivered to a site where enzymatic conversion of ethanolto acetate is desired. Possible sites include the mucosal membrane ofthe mouth, nasopharyngeal region, or rectum. This approach would not beuseful for modifying the microbiome in the colon for several reasons.For example: first, Whitmore teaches to deliver oxygen and enzymes tosites that would not result in an increase in oxygen in the colon;second, administration to reduce excess ethanol would be for a shortterm, whereas administration to modify the microbiome in the colontypically occurs over several days or weeks or months in order torealize a therapeutic benefit, such as weight management.

This invention meets needs in the art by providing pharmaceuticalpreparations that may be administered to a subject and that act in thecolon of the subject to increase the oxygen tension and/or redoxpotential and/or pH of the colon, thus changing the conditions in whichthe gut microbiome of the colon exists. A hypothesis of this inventionis that these endogenous changes to the environment of the colon will inturn change the relative abundance of various bacteria present in thecolon in a manner that mimicks, at least partially, changes to the gutmicrobiome previously observed in RYGB. Based in part on this discovery,the inventor provides herein methods comprising administering to asubject at least one agent that increases oxygen tension and/or redoxpotential and/or pH in the colon of a subject. In some embodiments theat least one agent is administered by administering an effective amountof the at least one agent to the small intestine and/or large intestineof the subject. Also provided are methods of treating a subject. In someembodiments the methods comprise providing a subject having or at riskof developing at least one condition selected from a metabolic diseaseor disorder, metabolic syndrome, cardiovascular disease, and excessweight; and administering a pharmaceutical composition of the inventionto the subject, to thereby treat the at least one condition selectedfrom a metabolic disease or disorder, metabolic syndrome, cardiovasculardisease, and excess weight in the subject. These and other features andaspects of the invention are disclosed more fully herein.

B. Definitions

As used herein, “subject” means any mammal. In some embodiments thesubject is a human. In some embodiments the subject is a primate. Insome embodiments the subject is a non-human mammal, such as a non-humanprimate. In some embodiments the subject is a farm animal or livestock.In some embodiments the subject is a pet or companion animal.

As used herein, an “active agent” is a chemical entity that acts toincrease the oxygen tension and/or redox potential and/or pH of anaqueous solution that approximates the conditions of a mammalian smallintestine or large intestine. Such an agent is variously referred toherein as an “active agent” and/or as an “agent that increases oxygentension and/or redox potential and/or pH in the colon of the subject tothereby manage the weight of the subject.”

As used herein, an “agent that increases oxygen tension and/or redoxpotential and/or pH in the colon of the subject to thereby manage theweight of the subject” is an agent that has at least one propertyselected from (1) increases the oxygen tension of a solution ofphosphate buffered saline (PBS), (2) increases redox potential of PBS,and (3) increases pH of PBS.

Whether an agent increases the oxygen tension of PBS is measured in thefollowing assay. A solution of phosphate buffered saline is provided.The PBS is 0.01 M phosphate buffered saline (NaCl 0.138 M; KCl—0.0027M); pH 7.4, at 25° C. The test agent is added to the PBS in differentconcentrations ranging from 1 nano molar to 1 molar. The solution isthen incubated at 37 C, and stirred at 20 rpm. Across titration withinthis concentration range, if the test agent increases oxygen tension byat least a predetermined cutoff, then the test agent is identified as anactive agent. Oxygen concentration is then measured using a commerciallyavailable dissolved oxygen meter. In some embodiments a test agent isidentified as an active agent if the test agent increases the oxygentension by at least 0.01 mg/L. In some embodiments a test agent isidentified as an active agent if the test agent increases the oxygentension by at least 0.02 mg/L. In some embodiments a test agent isidentified as an active agent if the test agent increases the oxygentension by at least 0.05 mg/L. In some embodiments a test agent isidentified as an active agent if the test agent increases the oxygentension by at least 0.10 mg/L. In some embodiments a test agent isidentified as an active agent if the test agent increases the oxygentension by at least 0.20 mg/L. In a preferred embodiment, the activeagent increases the oxygen tension by at least 0.10 mg/L.

Whether an agent increases the redox potential of PBS is measured in thefollowing assay. A solution of phosphate buffered saline is provided.The PBS is 0.01 M phosphate buffered saline (NaCl 0.138 M; KCl—0.0027M); pH 7.4, at 25° C. The test agent is added to the PBS in differentconcentrations ranging from 1 nano molar to 1 molar. The solution isthen incubated at 37 C, and stirred at 20 rpm. Across titration withinthis concentration range, if the test agent increases redox potential byat least a predetermined cutoff, then the test agent is identified as anactive agent. Redox potential is then measured using a commerciallyavailable redox meter. In some embodiments a test agent is identified asan active agent if the test agent increases the redox potential by atleast 0.1 mV. In some embodiments a test agent is identified as anactive agent if the test agent increases the redox potential by at least0.5 mV. In some embodiments a test agent is identified as an activeagent if the test agent increases the redox potential by at least 1.0mV. In some embodiments a test agent is identified as an active agent ifthe test agent increases the redox potential by at least 5.0 mV. In someembodiments a test agent is identified as an active agent if the testagent increases the redox potential by at least 10.0 mV. In someembodiments a test agent is identified as an active agent if the testagent increases the redox potential by at least 20.0 mV. In a preferredembodiment the active agent increases the redox potential by at least10.0 mV.

Whether an agent increases the pH of PBS is measured in the followingassay. A solution of phosphate buffered saline is provided. The PBS is0.01 M phosphate buffered saline (NaCl 0.138 M; KCl—0.0027 M); pH 7.4,at 25° C. The test agent is added to the PBS in different concentrationsranging from 1 nano molar to 1 molar. The solution is then incubated at37 C, and stirred at 20 rpm. Across titration within this concentrationrange, if the test agent increases redox potential by at least apredetermined cutoff, then the test agent is identified as an activeagent. The pH of the solution is then measured using a commerciallyavailable pH meter. In some embodiments a test agent is identified as anactive agent if the test agent increases the pH by at least 0.01 units.In some embodiments a test agent is identified as an active agent if thetest agent increases the pH by at least 0.02 units. In some embodimentsa test agent is identified as an active agent if the test agentincreases the pH by at least 0.05 units. In some embodiments a testagent is identified as an active agent if the test agent increases thepH by at least 0.10 units. In some embodiments a test agent isidentified as an active agent if the test agent increases the pH by atleast 0.20 units. In a preferred embodiment the active agent increasesthe pH by at least 0.10 units.

A skilled artisan will appreciate that a single agent may have any twoof these three activities or may have all three. That is, that a singleagent may increase oxygen tension and also increase redox potential; orthat a single agent may increase oxygen tension and also increase pH; orthat a single agent may increase redox potential and also increase pH;or that a single agent may increase oxygen tension, and increase redoxpotential, and increase pH. Thus, in some instances an agent that isidentified herein as an agent that increases one of oxygen tension,redox potential, and pH, that agent may also be an agent that increasesat least one additional feature selected from oxygen tension, redoxpotential, and pH. A non-limiting example is sodium percarbonate, whichincreases all of oxygen tension, redox potential and pH.

While exemplary active agents are disclosed herein the disclosed agentsare not intended to be limiting.

As used herein “condition” encompasses a disease, condition, ordisorder.

The term “effective amount” refers to the amount of active agent thatelicits a biological or medicinal response in a subject, including in atissue or system of the subject, and that is being sought by aresearcher, veterinarian, medical doctor, nutritionist, or otherclinician or caregiver or by a subject, which includes one or more ofthe following:

(1) Preventing the condition, for example, preventing a condition in asubject that may be predisposed to the condition but does not yetexperience or display the pathology or symptomatology of the condition;

(2) Inhibiting the condition, for example, inhibiting a condition in anindividual that is experiencing or displaying the pathology orsymptomatology of the condition (i.e., arresting further development ofthe pathology and/or symptomatology); and

(3) Ameliorating the condition, for example, ameliorating a condition inan individual that is experiencing or displaying the pathology orsymptomatology of the condition, (i.e., reversing the pathology and/orsymptomatology).

As used herein, an “increase in the relative abundance” of a group ofbacteria in the gut microbiota of a subject means an increase in theproportion of bacterial cells belonging to that group among the totalbacterial cells measured in an assay that samples the bacteria in themicrobiota of the gut of a subject. Relative abundance is defined as thenumber of bacteria belonging to that group divided by the total numberof bacteria measured. For example, if 20% of taxonomic markers in thegut microbiota of a subject obtained by 16S rRNA sequencing belong to agiven group of bacteria, the relative abundance of this group ofbacteria is scored as 20%. In other words, there would be 20 bacteriabelonging to this group in every 100 bacteria measured by sequencing oranother equivalent method. Maximum relative abundance is 100% and theminimum is 0%.

Whether someone is classified as overweight or obese is generallydetermined on the basis of their body mass index (BMI) which iscalculated by dividing body weight (kg) by height squared (m²). Thus,the units of BMI are kg/m² and it is possible to calculate the BMI rangeassociated with minimum mortality in each decade of life. As usedherein, “overweight” is defined as a BMI in the range 25-30 kg/m², and“obesity” or “obese” as a BMI greater than 30 kg/m².

As used herein the term “metabolic disorder”, refers to disorders,diseases, and conditions that are caused or characterized by abnormalweight gain, energy use or consumption, altered responses to ingested orendogenous nutrients, energy sources, hormones or other signalingmolecules within the body or altered metabolism of carbohydrates,lipids, proteins, nucleic acids or a combination thereof. A metabolicdisorder is associated with either a deficiency or excess in a metabolicpathway resulting in an imbalance in metabolism of nucleic acids,proteins, lipids, and/or carbohydrates. Factors affecting metabolisminclude, and are not limited to, the endocrine (hormonal) control system(e.g., the insulin pathway, the enteroendocrine hormones includingGLP-1, PYY or the like), the neural control system (e.g., GLP-1 or otherneurotransmitters or regulatory proteins in the brain) or the like. Somenon-limiting examples can be obesity, diabetes, including type IIdiabetes, insulin-deficiency, insulin-resistance, insulin-resistancerelated disorders, glucose intolerance, syndrome X, inflammatory andimmune disorders, osteoarthritis, dyslipidemia, metabolic syndrome,non-alcoholic fatty liver, abnormal lipid metabolism, cancer,neurodegenerative disorders, sleep apnea, hypertension, highcholesterol, atherogenic dyslipidemia, hyperlipidemic conditions such asatherosclerosis, hypercholesterolemia, and other coronary arterydiseases in mammals, and other disorders of metabolism. In someembodiments the metabolic disorder is diagnosed by a physician.

As used herein the term “treat,” “treating” or “treatment” refers to theadministration of therapy to an individual who already manifests atleast one symptom of a condition, or who has previously manifested atleast one symptom of a condition, or who is identified as at risk ofdeveloping a condition. For example, “treating” can include alleviating,abating or ameliorating a condition's symptoms, preventing additionalsymptoms, ameliorating or preventing the underlying metabolic causes ofsymptoms, inhibiting the condition, e.g., arresting the development ofthe condition, relieving the condition, causing regression of thecondition, relieving a second condition caused by the first condition,or stopping the symptoms of the condition either prophylacticly and/ortherapeutically. For example, the term “treating” in reference to acondition includes a reduction in severity of one or more symptomsassociated with a particular condition. Therefore, treating a conditiondoes not necessarily mean a reduction in severity of all symptomsassociated with a condition and does not necessarily mean a completereduction in the severity of one or more symptoms associated with acondition. For example, a method for treatment of cardiovascular diseasecan result in a reduction in blood pressure; however, the reduction inblood pressure does not need to be enough such that the individual has afully normal cardiovascular health. It has been shown that even modestdecreases in weight or related parameters such as BMI, waistcircumference and percent body fat, can result in improvement of health,for example, lower blood pressure, improved blood lipid profiles, or areduction in sleep apnea. For example, the term “treating” also includesreducing the rate of increase in severity in a condition alreadymanifested in a subject and/or reducing the rate of occurrence of newrelated conditions in the subject.

As used herein, “providing a subject having or at risk of developing atleast one condition” refers to a judgment made by a researcher,veterinarian, medical doctor, nutritionist, or other clinician orcaregiver, or by a subject, that a subject requires or will benefit ormay benefit from treatment.

As used herein, “weight management” means at least one of weight loss,maintenance of weight, maintenance of weight loss (also called weightmaintenance herein), controlling weight gain, body mass index (BMI)reduction, maintenance of BMI, maintenance of BMI reduction, andcontrolling BMI gain in a subject. In some embodiments the subject isoverweight or obese. In some embodiments the subject has at least oneweight-related condition. In some embodiments the at least oneweight-related condition is selected from a metabolic disease ordisorder, metabolic syndrome, and cardiovascular disease. In someembodiments the at least one weight-related condition is selected fromhypertension, dyslipidemia, and type 2 diabetes. In some embodiments atleast one symptom of the at least one weight-related condition isameliorated. For example, weight management for an individual who isoverweight or obese can mean losing weight with the goal of keepingweight in a healthier range. Also, for example, weight management for anindividual who is overweight or obese can include losing body fat(reduction in BMI) or circumference around the waist with or without theloss of body weight. Maintenance of weight loss (weight maintenance)includes preventing, reducing or controlling weight gain after weightloss. It is well known that weight gain often occurs after weight loss.Weight loss can occur, for example, from dieting, exercising, illness,drug treatment, surgery or any combination of these methods, but oftenan individual that has lost weight will regain some or all of the lostweight. Therefore, weight maintenance in an individual who has lostweight can include preventing weight gain after weight loss, reducingthe amount of weigh gained after weight loss, controlling weight gainafter weight loss or slowing the rate of weight gain after weight loss.As used herein, “weight management in an individual in need thereof”refers to a judgment made by a healthcare practitioner that anindividual requires or will benefit from weight management treatment.This judgment is made based on a variety of factors that are in therealm of a healthcare practitioner's expertise, but that includes theknowledge that the individual has a condition that is treatable by themethods disclosed herein. In some embodiments weight managementcomprises decreasing appetite. In some embodiments weight managementcomprises decreasing hunger.

As used herein, “aerobic bacteria” are bacteria that need oxygen togrow. Included within the aerobic bacteria are microaerophiles, whichare bacteria that require oxygen for energy production but are harmed byatmospheric concentrations of oxygen.

As used herein, “facultative anaerobes” are bacteria or othermicroorganisms such as fungi that make ATP by aerobic respiration ifoxygen is present, but are capable of switching to fermentation oranaerobic respiration if oxygen is absent.

As used herein, “sustained release” refers to a pharmaceuticalformulation containing a composition comprising at least one activeagent and at least one release rate modifier. The pharmaceuticalformulation is formulated so that the at least one release rate modifierreduces the rate of release of the at least one active agent incomparison to a similar formulation that does not contain at least onerelease rate modifier. The pharmaceutical formulation may also bereferred to as “formulated for sustained release” or as a “sustainedrelease formulation.” The pharmaceutical formulation may alternativelybe referred to as “formulated for extended release” or as an “extendedrelease formulation,” making reference to the reduced rate of releasethat extends the period of release.

As used herein, “delayed release” refers to a pharmaceutical formulationcontaining a composition comprising at least one active agent and acomposition coating that encases the at least one active agent andpreventing its immediate release in the stomach following oraladministration to a subject. The coating may be referred to as a“delayed release” coating. Generally the coating prevents the activeagent from interacting chemically with components of the stomach suchthat the active agent is protected from the stomach environment and isonly released from the coating once the microparticles reach the smallintestine and/or colon. Examples of delayed release coatings are entericcoatings well known in the art. An alternative type of delayed releasecoating is a coating that is digested by bacterial enzymes present inthe colon and absent from the stomach and/or the stomach and smallintestine. Enteric coatings are well known in the art. In general, anenteric coating is a polymer barrier that prevents the dissolution ordisintegration of an active agent in the gastric environment.

In some embodiments the period of release of active agent from asustained release formulation is increased in comparison to a comparableformulation that does not comprise the at least one release ratemodifier by a period of 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5hours, 6 hours, 12 hours, or 24 hours. In some embodiments the period ofrelease of active agent from a sustained release formulation isincreased in comparison to a comparable formulation that does notcomprise the at least one release rate modifier by a period of at least0.5 hours, at least 1 hour, at least 2 hours, at least 3 hours, at least4 hours, at least 5 hours, at least 6 hours, at least 12 hours, or atleast 24 hours.

Throughout this specification, unless specifically stated otherwise orthe context requires otherwise, reference to a single step, compositionof matter, group of steps or group of compositions of matter shall betaken to encompass one and a plurality (i.e. one or more) of thosesteps, compositions of matter, groups of steps or group of compositionsof matter.

Each embodiment described herein is to be applied mutatis mutandis toeach and every other embodiment unless specifically stated otherwise.

Those skilled in the art will appreciate that the invention(s) describedherein is susceptible to variations and modifications other than thosespecifically described. It is to be understood that the invention(s)includes all such variations and modifications. The invention(s) alsoincludes all of the steps, features, compositions and compounds referredto or indicated in this specification, individually or collectively, andany and all combinations or any two or more of said steps or featuresunless specifically stated otherwise.

The present invention(s) is not to be limited in scope by the specificembodiments described herein, which are intended for the purpose ofexemplification only. Functionally-equivalent products, compositions andmethods are clearly within the scope of the invention(s), as describedherein.

It is appreciated that certain features of the invention(s), which are,for clarity, described in the context of separate embodiments, can alsobe provided in combination in a single embodiment. Conversely, variousfeatures of the invention(s), which are, for brevity, described in thecontext of a single embodiment, can also be provided separately or inany suitable subcombination.

C. Agents that Increase Oxygen Tension and/or Redox Potential and/or pH

A skilled artisan will appreciate that any agent that increases oxygentension and/or redox potential and/or pH (i.e., any active agent) may beused in the pharmaceutical preparations and methods of this invention.

Agents that increase oxygen tension include agents that carry oxygen tothe large intestine and agents that increase production of oxygen fromstores in the gut and/or from added oxygen stores. Any oxygen carriermay be used to increase oxygen tension, including a molecule that reactsto release oxygen. For example, in some embodiments the at least oneagent comprises a peroxide functional group. A peroxide functional groupis a group comprising or consisting of an O—O single bond or theperoxide anion. In contrast to oxide ions, the oxygen atoms in theperoxide ion have an oxidation state of −1. The simplest stable peroxideis hydrogen peroxide. In some embodiments at least one agent is hydrogenperoxide. In some embodiments the at least one peroxide is an organicperoxide. In some embodiments the at least one peroxide is an organicperoxide selected from acetyl acetone peroxide, acetyl benzoyl peroxide,ascaridole, benzoyl peroxide, di-(1-naphthoyl)peroxide, diacetylperoxide, ethyl hydroperoxide, ergesterol peroxide, iodoxy compounds,methyl isobutyl ketone peroxide. In some embodiments the at least oneagent is an inorganic peroxide. In some embodiments the at least oneagent is an inorganic peroxide selected from Ammonium persulfate,Calcium peroxide, Magnesium peroxide, Potassium persulfate, Sodiumperoxide, Lithium peroxide, Barium peroxide and Sodium perborate. Insome embodiments the at least one agent is carbamide peroxide. In someembodiments the at least one agent is Sodium percarbonate.

In some embodiments, sodium percarbonate is formulated as granules. Insome embodiments sodium percarbonate granules are coated with protectivecoating or is chosen from commercially available coated particles suchas FB 400C, FB 700C, OXYPER (Solvay), Provox-C(OCI corporation),ECOX-C(Kemira). The coating can be based on sodium carbonate, sodiumchloride and sodium metasilicate or combinations thereof. Coating may beused to improve the stability of the final product.

In some embodiments, agents that increase oxygen tension are selectedfrom reactive oxygen species (ROS) other than peroxides. Reactive oxygenspecies in the presence of catalysts or corresponding enzymes ofmammalian or bacteria can be converted or decomposed into hydrogenperoxide or oxygen. In some embodiments one such agent is selected fromsuperoxides, dioxygenyls, ozones and ozonides. In some embodiments onesuch agent is selected from singlet oxygen, hydroxyl radical,superoxide, nitric oxide, ozone, peroxyl, lipid peroxyl, hypochloricacid. In some embodiments, agents that increase oxygen tension areselected from reactive nitrogen species such as nitric oxide which candecompose into nitrogen and oxygen gas. In some embodiments, such agentsare selected from nitrous oxide, peroxynitrite, peroxynitrous acid,nitrosoperoxycarbonate, dinitrogen trioxide, nitroxyl anion, nitrogendioxide, nitrous acid, nitrousyl cation, nitryl chloride, nitrosoniumcation, higher oxides of nitrogen, S-nitrosothiols, and dinitrosyl ironcomplexes.

In some embodiments, the agent that increases oxygen tension is anoxygen carrier. In some embodiments the oxygen carrier is aperfluorocarbon. In some embodiments the perfluorocarbon is selectedfrom perfluorodecalin, bromoperfluoro-n-octane (perfluorobron),dichloroperfluoro-n-octane, triperfluoropropylamine. Oxygen gas may bedissolved in perfluorocarbon fluids and delivered to the colon withenteric coating or timed release technology. In some embodiments, theoxygen carrier is microbubble preparation of oxygen gas created byencapsulation of oxygen gas into polymetric bubbles or lipid based ofemulsion of air/oxygen gas. In some embodiments, oxygen gas may bedirectly delivered to the gut to increase oxygen tension. In someembodiments, oxygen gas is encapsulated into enterically coated capsulesand delivered to increase oxygen tension in the colon using pH or timedrelease formulations.

Agents that increase production of oxygen from stores in the gut and/orfrom added oxygen stores include any enzyme that catalyzes a chemicalreaction that produces oxygen. An example is catalase, a common enzymefound in nearly all living organisms exposed to oxygen. Catalasecatalyzes the decomposition of hydrogen peroxide to water and oxygen.Accordingly, in some embodiments the agent that increases oxygen tensionis a catalase. In some embodiments the catalase is a human catalase. Insome embodiments the catalase is a non-human catalase. In someembodiments the catalase is a mammalian catalase. In some embodimentsthe catalase is a non-mammalian catalase. In some embodiments thecatalase is used as the only agent that increases oxygen tension. Insome embodiments the catalase is a plant catalase such as potatocatalase. In some embodiments the catalase is liver catalase, such asbovine catalase. In some embodiments the catalase is blood catalase. Insome embodiments the catalase is bacterial catalase. In some embodimentsthe catalase is obtained from Micrococcus genus. In some embodiments thecatalase is fungal catalase. In some embodiments the catalase isobtained from Aspergillus niger. In some embodiments the catalase isrecombinant. In such embodiments catalase increases oxygen tension byincreasing the rate of decomposition of endogenous hydrogen peroxide towater and oxygen. In other embodiments catalase and hydrogen peroxideare used together.

Additional examples of agents that catalyze a chemical reaction thatproduces oxygen include additional peroxide catalysts. In someembodiments the peroxide catalyst is an inorganic peroxide catalyst. Insome embodiments the inorganic peroxide catalyst is selected frommanganese(IV) oxide (manganese dioxide, MnO2), lead(IV) oxide (leaddioxide, PbO2), iron(III) oxide (red iron oxide, Fe2O3), silver nitrateand potassium iodide. In some embodiments the peroxide catalyst is anorganic peroxide catalyst. In some embodiments the organic peroxidecatalyst is selected from blood and bacteria. In some embodiments theorganic peroxide catalyst is a peroxidase. Peroxidases are a largefamily of enzymes that typically catalyze a reaction of the form:ROOR′+electron donor (2 e−)+2H+→ROH+R′OH

In some embodiments the at least one agent is an agent that increasespH. Non-limiting examples of agents that increase pH include bicarbonatesalts, carbonate salts, bases, and buffers. In some embodiments theagent that increases pH is bicarbonate. Bicarbonate acts physiologicallyto regulate pH in the small intestine. It is released from the pancreasin response to the hormone secretin to neutralize the acidic chymeentering the duodenum from the stomach. In some embodiments the at leastone agent is bicarbonate. In some embodiments the at least one agentcomprises a bicarbonate group. Examples include bicarbonate salts, suchas sodium bicarbonate, potassium bicarbonate, caesium bicarbonate,magnesium bicarbonate, calcium bicarbonate, ammonium bicarbonate. Insome embodiments the at least one agent is a bicarbonate salt. In someembodiments the bicarbonate salt is selected from Sodium HydrogenCarbonate, Beryllium Hydrogen Carbonate, Magnesium Hydrogen Carbonate,Calcium Hydrogen Carbonate, Potassium Hydrogen Carbonate, BariumHydrogen Carbonate, Copper(II) Hydrogen Carbonate, Iron(III) HydrogenCarbonate, Aluminum Hydrogen Carbonate, Lithium Hydrogen Carbonate,Lead(II) Hydrogen Carbonate, Tin(IV) Hydrogen Carbonate, Iron(II)Hydrogen Carbonate, Ammonium Hydrogen Carbonate, Mercury(II) HydrogenCarbonate, Lead(IV) Hydrogen Carbonate, Manganese(II) HydrogenCarbonate, Cesium Hydrogen Carbonate, Silver Hydrogen Carbonate, TinHydrogen Carbonate, Copper(I) Hydrogen Carbonate, Zinc HydrogenCarbonate, Rubidium Hydrogen Carbonate, Nickel(II) Hydrogen Carbonate,Cadmium Hydrogen Carbonate, Strontium(II) Hydrogen Carbonate, Mercury(I)Hydrogen Carbonate, Chromium(II) Hydrogen Carbonate, Gold(III) HydrogenCarbonate, Cobalt(II) Hydrogen Carbonate, and Cobalt(III) HydrogenCarbonate.

In some embodiments the at least one agent is a carbonate. In someembodiments the carbonate is a carbonate salt. In some embodiments thecarbonate salt is selected from Sodium Carbonate, Sodium percarbonate(Sodium Carbonate Peroxide), Calcium Carbonate, Cobalt(III) Carbonate,Copper(I) Carbonate, Potassium Carbonate, Ammonium Carbonate,Chromium(III) Carbonate, Iron(III) Carbonate, Aluminum Carbonate,Tin(IV) Carbonate, Lead(IV) Carbonate, Magnesium Carbonate, Iron(II)Carbonate, Tin(II) Carbonate, Chromium(VI) Carbonate, Silver(I)Carbonate, Titanium(IV) Carbonate, Vanadium(III) Carbonate, Copper(II)Carbonate, Zinc(II) Carbonate, Lithium Carbonate (Lithium Salt),Cobalt(II) Carbonate, Nickel(III) Carbonate, Sodium CarbonateDecahydrate, Mercury(I) Carbonate, Barium Carbonate, Lead Carbonate,Mercury(II) Carbonate, Chromium(II) Carbonate, Strontium Carbonate, andVanadium(V) Carbonate.

In some embodiments the at least one agent is a base. In someembodiments the base is a strong base, such as a base selected fromSodium Hydroxide, Aluminum Hydroxide, Calcium Hydroxide, BariumHydroxide, Magnesium Hydroxide, Iron(III) Hydroxide, Ammonium Hydroxide,Potassium Hydroxide, Chromium(III) Hydroxide, Zinc Hydroxide, Lead(II)Hydroxide, Platinum(IV) Hydroxide, Vanadium(V) Hydroxide, BerylliumHydroxide, Copper(II) Hydroxide, Lead(IV) Hydroxide, Vanadium(III)Hydroxide, Iron(II) Hydroxide, Nickel(II) Hydroxide, Tin(IV) Hydroxide,Silver Hydroxide, Strontium Hydroxide, Tin(II) Hydroxide, LithiumHydroxide, Manganese(II) Hydroxide, Chromium(II) Hydroxide, NickelOxo-hydroxide, Mercury(II) Hydroxide, Cadmium Hydroxide, and Copper(I)Hydroxide, Manganese(IV) Hydroxide. In some embodiments the base is asuperbase, such as a base selected from Butyl lithium (n-C4H9Li),Lithium diisopropylamide (LDA) [(CH3)2CH]2Nli, Lithium diethylamide(LDEA) (C2H5)2Nli, Sodium amide (NaNH2), Sodium hydride (NaH), andLithium bis(trimethylsilyl)amide [(CH3)3Si2NLi. In some embodiments thebase is an organic base, such as a base selected from Amines,Nitrogen-containing heterocyclic compounds, Urea, Ammonia, pyridine,methyl amine, imidazole, benzimidazole, histidine, and andphosphazene.In some embodiments the agent is a weak base.

In some embodiments the agent is a buffer, such as a buffer selectedfrom MES, Bis-Tris, ADA, aces, PIPES, MOPSO, Bis-Tris Propane, BES,MOPS, TES, HEPES, DIPSO, MOBS, TAPSO, Trizma, HEPPSO, POPSO, TEA, EPPS,Tricine, Gly-Gly, Bicine, HEPBS, TAPS, AMPD, TABS, AMPSO, CHES, CAPSO,AMP, CAPS, and CABS. In some embodiments the buffer is a CitricAcid—Na2HPO4 Buffer Solution. In some embodiments the buffer is aNa2HPO4-NaH2PO4 Buffer Solution. In some embodiments the buffer is aSodium Carbonate—Sodium Bicarbonate Buffer Solution. In some embodimentsthe buffer is a Imidazole (glyoxaline)—HCl buffer solution.

In some embodiments the at least one agent is an agent that increasesredox potential. In some embodiments, the agent that increases redoxpotential is an oxidizing agent. In some embodiments, the oxidizingagent is selected from compounds containing halogens such as Fluorine(F), Chlorine (Cl), Bromine (Br), Iodine (I), and Astatine (At). In someembodiments, the oxidizing agent is selected from Aluminium nitrate,Ammonium chlorate, Ammonium dichromate, Ammonium nitrate, Ammoniumnitrite, Ammonium perchlorate, Ammonium permanganate, Ammoniumpersulfate, Antimony pentachloride, Barium chlorate, Barium chromate,Barium manganate, Barium nitrate, Barium perchlorate, Barium peroxide,Benedict's reagent, Bismuth pentafluoride, Bromic acid, Bromine, Brominemonochloride, Bromine pentafluoride, Bromine trifluoride, Bromus acid,Cadmium nitrate, Caesium chromate, Caesium nitrate, Caesium perchlorate,Calcium bromate, Calcium chlorate, Calcium chromate, Calciumhypochlorite, Calcium iodate, Calcium nitrate, Calcium permanganate,Calcium peroxide, Ceric ammonium nitrate, Cerium(III) methanesulfonate,Chloric acid, Chlorine, Chlorine monofluoride, Chlorine nitrate,Chlorine pentafluoride, Chlorine trifluoride, Meta-Chloroperoxybenzoicacid, N-Chlorosuccinimide, Chlorous acid, Chromate and dichromate,Chromic acid, Chromium nitrate, Chromyl chloride, Chromyl fluoride,Cobalt(II) chlorate, Cobalt(II) nitrate, Collins reagent, Copper(II)acetate, Copper(II) hydroxide, Copper(II) nitrate, Copper(II)perchlorate, Dess-Martin periodinane, Dichlorine heptoxide, Dinitrogentetroxide, Dioxygen difluoride, Fehling's solution, Fenton's reagent,Fluorine, Fluorine perchlorate, Gadolinium(III) nitrate, Hill reagent,Hydrazine nitrate, Hydrogen peroxide, Hydrogen peroxide-urea,Hypobromous acid, Hypochlorous acid, Hypoiodous acid, Iodic acid,Iodine, Iodine heptafluoride, Iodine monochloride, Iodine pentafluoride,Iodine pentoxide, Iodine trichloride-Iodobenzene dichloride Iodous acid,Iron(III) chromate, Iron(III) nitrate, Jones reagent, Lead(II) chromate,Lead(II) nitrate, Lead(IV) acetate-Lithium chlorate, Lithiumhypochlorite, Lithium nitrate, Lithium nitrite, Lithium perchlorate,Lithium peroxide, Magnesium monoperoxyphthalate, Magnesium nitrate,Magnesium perchlorate, Manganese(III) acetate, Mercury(II) nitrate,Nickel chromate, Nickel(II) nitrate, Nitronium perchlorate,Nitrosyl-O-hydroxide, Nitrous acid, Osmium tetroxide, Oxygen, Oxygendifluoride, Ozone, Palladium(II) nitrate, Perbromic acid, Perchlorate,Perchloric acid, Performic acid, Periodic acid, Permanganic acid, Peroxyacid, Peroxymonosulfuric acid, Potassium bromate, Potassium chromate,Potassium dichromate, Potassium hypochlorite, Potassium iodate,Potassium nitrate, Potassium nitrite, Potassium perchlorate, Potassiumperiodate, Potassium permanganate, Potassium peroxide, Potassiumperoxymonosulfate, Potassium persulfate, Potassium superoxide, Potassiumtetraperoxochromate(V), Potassium trioxochlorochromate, Pyridiniumchlorochromate, Rubidium nitrate, Rubidium perchlorate, Scandiumnitrate, Selenic acid Selenium hexasulfide, Selenium trioxide, Selenousacid, Silver bromate, Silver chlorate, Silver chromate, Silverdichromate, Silver iodate, Silver nitrate, Silver perchlorate, Singletoxygen, Sodium bromate, Sodium chlorate, Sodium chlorite, Sodiumchromate, Sodium dichromate, Sodium hypochlorite, Sodium fluoride,Sodium iodate, Sodium nitrate, Sodium nitrite, Sodium perborate, Sodiumpercarbonate, Sodium perchlorate, Sodium periodate, Sodium permanganate,Sodium peroxide, Sodium persulfate, Sodium superoxide, Stannousfluoride, Strontium bromate, Strontium chlorate, Strontium nitrate,Strontium peroxide, Sulfuric acid, Superoxidant, Telluric acid,Tetrapropylammonium perruthenate, Thallium(III) nitrate, Tollens'reagent, Trinitroethylorthocarbonate, Trinitroethylorthoformate, Uranylnitrate, Uranyl peroxide, Zinc chlorate, Zinc nitrate, and Zincperoxide. In some embodiments, the agent that increases redox potentialis a fluoride containing compound. In some embodiments, the agent thatincreases redox potential is selected from Sodium fluoride, Sodiummonofluorophosphate and Stennous fluoride.

D. Pharmaceutical Preparations

This invention provides pharmaceutical preparations comprising aplurality of particles, each particle comprising: 1) a core comprisingat least one agent that increases oxygen tension and/or redox potentialand/or pH in the colon of a subject following administration to thesubject, and 2) an enteric coating encasing the core.

This invention also provides pharmaceutical preparations comprising aplurality of particles and a capsule or sachet encasing the plurality ofparticles. In some embodiments the pharmaceutical preparation comprises:A) A plurality of particles, each particle comprising: 1) a corecomprising at least one agent that increases oxygen tension and/or redoxpotential and/or pH in the colon of a subject following administrationto the subject, and 2) an enteric coating encasing the core; and B) acapsule or sachet encasing the plurality of particles.

The pharmaceutical preparations of the invention typically comprise aplurality of particles, each particle comprising: 1) a core comprisingat least one agent that increases oxygen tension and/or redox potentialand/or pH in the colon of a subject following administration to thesubject, and 2) an enteric coating encasing the core. In someembodiments the cores of the plurality of particles have maximumdimensions of from 0.1 to 2 mm. In some embodiments the cores of theplurality of particles have maximum dimensions of from 0.2 to 1.0 mm. Insome embodiments the cores of the plurality of particles have maximumdimensions of from 0.4 to 1.2 mm. In some embodiments the cores of theplurality of particles have maximum dimensions of from 0.6 to 1.4 mm. Insome embodiments the cores of the plurality of particles have maximumdimensions of from 0.8 to 1.6 mm. In some embodiments the cores of theplurality of particles have maximum dimensions of from 1.0 to 1.8 mm. Insome embodiments the cores of the plurality of particles have maximumdimensions of from 0.4 to 0.8 mm. In some embodiments the plurality ofparticles have maximum dimension of no more than 3.0 mm. In someembodiments the plurality of particles have maximum dimension of no morethan 2.5 mm. In some embodiments the plurality of particles have maximumdimension of no more than 2.0 mm. In some embodiments the plurality ofparticles have maximum dimension of no more than 1.5 mm. In someembodiments the plurality of particles have maximum dimension of no morethan 1 mm. In some embodiments the plurality of particles have maximumdimensions of from 2.0 to 3.0 mm. In some embodiments the plurality ofparticles have maximum dimensions of from 1.5 to 2.5 mm. In someembodiments the plurality of particles have maximum dimensions of from1.0 to 2.0 mm. In some embodiments the plurality of particles havemaximum dimensions of from 2.5 to 3.0 mm. In some embodiments theplurality of particles have maximum dimensions of from 2.0 to 2.5 mm. Insome embodiments the plurality of particles have maximum dimensions offrom 1.5 to 2.0 mm. In some embodiments the plurality of particles havemaximum dimensions of from 1.0 to 1.5 mm.

In some embodiments the core consists of the active agent. In someembodiments the core consists of a plurality of active agents. In someembodiments the core comprises at least one active agent. In someembodiments the core comprises at least one active agent and at leastone release rate modifier. In some embodiments the core consists of atlease one active agent and the core is coated by a release ratemodifier.

The release rate modifier controls the rate of release of the at leastone active agent when the in the small intestine. In some embodiments,the release rate modifier is a polymer that is degraded by the gutmicrobiota selected from azopolymers, starches, dextrans,mucopolysaccharides (e.g. inulin, guar gum, pectin, chondroitin sulfate,alginic acid). In some embodiments the release rate modifier comprises ahydrophobic material, such as a waxy solid. Exemplary waxes suitable touse as the hydrophobic material include hydrocarbon waxes, such asparaffin wax and the like, which are substantially or entirely free ofunsaturation. Exemplary paraffin waxes are higher alkanes and mixturesof higher alkanes of the general formula C_(n)H2_(2n+2), wheretypically, 20<n<50, and thus have no unsaturation. They are solid atambient temperatures and melt-processable.

In some embodiments the release rate modifier comprises a hydrophilicmaterial. In some embodiments the hydrophilic material is a hydrophilicorganic polymer which is capable of hydrogen bonding and solid atambient temperatures (25° C.), hydrophilic and/or water soluble powders,and combinations thereof. In some embodiments the release rate modifiercomprises a hydrophilic material and a hydrophobic material. That is, itcomprises a plurality of materials and least one of the materials ismore hydrophilic than at least a second material (that is in turn morehydrophobic than the first material).

In some embodiments the hydrophilic material is dispersed in thehydrophobic material. In the case of organic polymers, the hydrophilicmaterial may be a material which is insoluble or substantially insolublein the hydrophobic material such that it forms discrete regions where itis of high concentration in the hydrophobic material (or forms aseparate). The regions may be spaced from each other by the hydrophobicmaterial. In the case of hydrophilic and/or water soluble powders, thepowder may be dispersed throughout the hydrophobic material, or in oneembodiment, more highly concentrated near an outer surface thereof.

In the case of hydrophilic and/or water soluble powders as release ratemodifiers, these may be present in a total concentration of from 0.001wt. % to 30 wt. %, such as 0.1-20 wt. %, or 1.0 to 10 wt. %. Examples ofhydrophilic powders include anhydrous inorganic particles, such assilicon dioxide, e.g., hydrophilic silica and silica nanopowders.Exemplary water-soluble powders include water-soluble acids and saltsthereof, such as anhydrous phosphate salts, e.g., sodium polyphosphate,sodium tripolyphosphate, sodium pyrophosphate; anhydrous citric acid andsalts thereof, such as alkali metals salts, e.g., sodium citrate;anhydrous sodium sulfate, anhydrous magnesium salts, such as magnesiumsulfate and magnesium chloride. Combinations of such release agents maybe employed. The hydrophilic and/or water soluble powders, such assilica, may have an average particle size of, for example, 1-100nanometers (nm), e.g., 5-20 nm, and a surface area of, for example50-400 m²/g. Hydrophilic fumed silica, for example, may be obtainedunder the tradename AEROSIL™ from Evonik Industries with a specificsurface area (measured by the BET method) in the range of 90-300 m²/g.As an example, AEROSIL™ 200 has a specific surface area of 200 m²/g.

When hydrophilic organic polymers are used as release rate modifiers,these may be present at a total concentration of from 0.5 wt. % to 40wt. %, e.g., 1-35 wt. %, or 10-30 wt. %. In one embodiment, thehydrophilic polymer has a melting point of at least 30° C. or at least40° C., such as up to 80° C. The hydrophilic polymer can have a weightaverage molecular weight of at least 300. Examples of suitablehydrophilic organic polymers include polyalkylene glycols, such aspolyethylene glycol and polypropylene glycol, and esters thereof,polyamide compounds (e.g., polyvinylpyrrolidone), poly(vinyl acetate),poly(vinyl alcohol), poly(acrylic acid), polyacrylamide,polyoxylglycerides, such as lauroyl, oleoyl, and stearoylpolyoxylglycerides, which are mixtures of monoesters, diesters, andtriesiers of glycerol and monoesters and diesters of polyethyleneglycols (e.g., lauroyl macrogolglycerides, such as GELUCIRE™ 44/14,available from Gattefosse, which has a melting point of 44° C. and anHLB of 14), and ethylene oxide derivatives thereof, poloxamers, whichare triblock copolymers having a central hydrophobic block ofpoly(propylene oxide) and two side blocks of poly(ethylene oxide) (e.g.,poloxamer 188, which has a melting point 52° C.), and derivativesthereof, and mixtures thereof.

Exemplary polyethylene glycols (PEG) suitable for the release ratemodifier may have a weight average molecular weight of from 300 daltonsto 50,000 daltons, such as about 600-35000, or 1000 to 5,000 daltons.Such materials are commercially available as PEG 1000 (melting point37-40° C.), PEG 1500 (melting point 44-48° C.), PEG 2000 (melting point49-52° C.), and the like. A combination of polyethylene glycols havingdifferent molecular weights may be employed to tailor the release rate.For example a mixture may be formed by combining, e.g., in a ratio offrom 1:10 to 10:1, a polyethylene glycol having a molecular weight ofabout 500-1200 (on average), such as PEG 1000, with a polyethyleneglycol having a molecular weight of at least 1500 or at least 1800 (onaverage), such as PEG 1500 or PEG 2000. In one embodiment, a combinationof PEGs with average molecular weight ranging from 300 daltons to 50,000daltons may be mixed on appropriate amounts to provide a mixture whichis liquid at a temperature of 35-70° C., such as 45-60° C. For example,PEG with an average molecular weight of 20,000 and PEG 1500 have meltingpoints of 60-65° C. and 44-48° C., respectively, and a mixture of PEG1500 and PEG 20,000 may be liquid at about 55° C., depending on theratio.

In the case of hydrophilic organic polymers, such as PEG, discreteregions in which the polymer is localized may have an average size of,for example, at least 0.1 or at least 0.5 nm, and can be up to 100 nm,or up to 20 nm, e.g., 0.5-5 nm. For example, the hydrodynamic radius ofglycerol is 0.3 nm and that of PEG 1000, PEG 2000 and PEG 4000 isapproximately 0.9, 1.4 and 1.9 nm, respectively.

A ratio of hydrophobic material to the hydrophilic material in therelease rate modifier may be from 1:99 to 99:1, expressed by weight,such as from 2:98 to 98:2, or from 10:90 to 90 10, or from 15:85 to85:15. The ratio can be at least 30:70, or at least 40:60, or at least60:40. For example, in the case of polymers, such as PEG, the ratio ofhydrophobic material to release rate modifier may be about 60:40 orabout 50:50. For hydrophilic and/or water soluble powders, the ratio ofhydrophobic material to the release rate modifier may be higher, such asat least 85:15, or at least 90:10.

In some embodiments, the hydrophilic material increases the rate ofrelease of the active ingredient, as compared with the hydrophobicmaterial alone. For example, the amount of active ingredient released(e.g., expressed as weight of hydrogen peroxide), may be at least 10%greater or at least 50% greater, over an initial period of two hours,than for the equivalent dosage form formed without the hydrophilicmaterial, when exposed to the same aqueous conditions (e.g., a bufferedrelease medium, at a temperature of 30-40° C.).

In some embodiments, the release rate modifier may provide a moreuniform rate of release of the active agent than equivalent dosage formsformed without the release rate modifier, when exposed to the sameaqueous conditions (e.g., buffered release medium at a temperature of30-40° C.). For example, the initial release rate (expressed as wt. ofactive agent/hr), over about two hours, may be, on average, less thanthat of an equivalent dosage form without the release rate modifier andmay be, on average, higher than that of equivalent dosage form in thesubsequent two hour period.

In some embodiments, the exemplary dosage form formed with the releaserate modifier release at least 10%, or at least 20%, or at least 30%, orat least 40%, or at least 50%, or at least 60%, or at least 70%, or atleast 80%, or at least 90% by weight of the total amount of active agentthat they contain over a period of 4 hours, 6 hours, or 8 hours aftercontact with the small intestine or aqueous medium at 30°-40° C. In someembodiments, the exemplary dosage form formed with the release ratemodifier release less than 25% or less than 50% by weight of the totalamount of active agent that they contain over a period of 2 hours aftercontact with the small intestine or aqueous medium at 30°-40° C.

As will be appreciated from the foregoing, the amount and type ofrelease rate modifier can be selected to tailor the release rateaccording to the desired application.

In some embodiments, the release rate modifier further includes anemulsifier, dispersed in the hydrophobic material. Exemplary nonionicsurfactants suitable as emulsifiers include fatty acids, polyol fattyacid esters, such as polyglycerol esters, fatty alcohol polyglycolethers, alkylphenol polyglycol ethers, fatty acid polyglycol esters,fatty acid amide polyglycol ethers, fatty amine polyglycol ethers,alkoxylated triglycerides, mixed ethers and mixed formals, optionallypartly oxidized alk(en)yl oligoglycosides or glucuronic acidderivatives, fatty acid-N-alkyl glucamides, protein hydrolyzates(particularly wheat-based vegetable products), sugar esters, sorbitanesters, polysorbates, amine oxides and combinations thereof. As examplesof suitable emulsifiers, nonionic surfactants with a lowhydrophile-lipophile balance (HLB) may be used. The HLB may be from 2-5.Surfactants that are able to form micelles are able to improve thestability of hydrogen peroxide. Examples of these emulsifiers includeC12-C24 fatty acids, such as lauric acid (CI 2), myristic acid (C14),palmitic acid (C16), stearic acid (C18), oleic acid (C18), linoleic acid(CI 8), and mixtures thereof. Such fatty acid emulsifiers can beobtained from Sigma-Aldrich under the tradename SPAN™, such as SPAN™ 60,which has an HLB of 4.7, SPAN™ 65, with an HLB of 2.1, SPAN™ 80, with anHLB of 4.3. Exemplary polyglycerol esters include polyglycerolpolyricinoleate (PGPR), which has an HLB of 3, and is available fromEvronik Industries, Essen Germany, or Danisco. A blend of surfactantshaving a high HLB and low HLB value may be used.

In some embodiments the pharmaceutical preparation comprises from 10 to1000 particles, from 10 to 100, from 100 to 200, from 200 to 300, from300 to 400, from 400 to 500, from 500 to 600, from 600 to 700, from 700to 800, from 800 to 900, or from 900 to 1000 particles. In someembodiments the pharmaceutical preparation comprises from 50 to 100,from 50 to 200 or from 100 to 500 particles.

A delayed release coating is applied to the core to form particles inorder to delay the release of active ingredients until the particle haspassed through the acidic environment of the stomach and has reached thesmall intestine and/or colon. The physical chemical environment of thestomach and gastric physiology are highly variable, subject to multiplefactors such as disease state, medication, age, and eating. For examplein the fasted state stomach, the pH is less than 2 in healthyindividuals, and gastric emptying occurs approximately every 30 minutes.However in the fed state (immediately after a meal), gastric emptying isdelayed for 2 to 4 hours and gastric pH can be as high as pH 4.

For these reasons, in some embodiments the delayed release coatingsystem is flexible. In some embodiments the pharmaceutical preparationis recommended to be taken on an empty stomach. In that case the delayedrelease coating may be resistant to the acidic stomach environment for arelatively short time and would not be expected to be subjected tostrong mechanical attrition in the stomach. On the other hand, to allowfor possible ingestion in the fed state, or where subsequent releasefrom the intestine is not intended to be immediate, the coating maytypically be sufficiently robust to withstand prolonged attrition in thestomach or to generally release more slowly in the alkaline environment.

The delayed release coating is typically (but not always) an entericcoating. There is a long history of use of enteric coatings on tabletsand smaller multi-particulate dosage forms in the pharmaceuticalindustry. Generally polymers with acidic functional groups are chosenfor enteric coatings. In the acid environment of the stomach these acidgroups of the polymers are un-ionized, thus rendering the polymer waterinsoluble. However in the more neutral and alkaline pH of the intestine(pH 6.8-7.2), the functional groups ionize and the polymer film coatingbecomes water soluble.

Examples of enteric film coatings that may be used in embodiments of theinvention include methacrylic acid copolymers, polyvinyl acetatephthalate, cellulose acetate phthalate, hydroxypropyl methylcellulosephthalate and hydroxypropyl methylcellulose acetylsuccinate. In someembodiments these water soluble coatings are applied from organicsolvent based coating solutions. In some embodiments an aqueous baseddispersion and/or pseudo-latex system of comprising at least one of theabove polymers is used. In some embodiments the polymers used forenteric coating are found in the Food Chemicals Codex (FCC), have directfood additive status, and/or have generally regarded as safe (GRAS)status. In some embodiments, commercially available enteric coatingssuch as EUDAGRIT™ coatings (Evonik Industries) are used. In someembodiments, EUDAGRIT™ enteric coatings for dissolution at specific pHlevels such as pH above 5.5 (Eudragit® L30D-55), pH above 5.6, pH above6.0, pH above 6.8 (Eudragit® FS 30D) and pH above 7.0 (Eudragit® S100)are used. In some embodiments, the core of the formulation includessolid powder of an anionic copolymer matrix based on methacrylic acidand methyl methacrylate such as Eudagrit S 100 in order to achieve pHdependent sustained release of actives in distal ileum and colon.EUDRAGIT® FS 30 D is the aqueous dispersion of an anionic copolymerbased on methyl acrylate, methyl methacrylate and methacrylic acid. Theratio of the free carboxyl groups to the ester groups is approx. 1:10.

Several strategies may be used to provide a food grade enteric coatingfor the pharmaceutical preparations of the invention. For example, anaqueous ethylcellulose (EC) based pseudo-latex may be used inconjunction with sodium alginate. For example, in some embodiments theNutrateric™ nutritional enteric coating system marketed by Colorcon Inc.of Westpoint, Pa. is used. This coating is supplied as a two componentsystem in the form of an aqueous ammoniated EC dispersion with 25%solids and a separate container of sodium alginate in powder form. Toprepare the final coating solution, the sodium alginate is firstdispersed and dissolved in water for 60 minutes and EC dispersion isthen added to the alginate solution, ensuring that the amount of waterused is appropriate to achieve a final recommended dispersed solidsconcentration of 10% by weight.

An alternative approach is the use of shellac on its own or incombination with other additives. Shellac is a natural, food approved,resinous material obtained from the exudate of the insect Karria lacca.It is a complex mixture of materials. The two main components withenteric properties being shelloic and aleuritic acid. Shellac may beused in the form of organic solvent based solutions. To obviate the useof solvents, neutralized aqueous shellac solutions are commerciallyavailable. EP 1 579 771 A1 describes a water based shellac dispersionwhich comprises shellac, a basic amino acid, a basic phosphate andwater. The basic amino acid being selected from the group consisting ofarginine, lysine and ornithine. Several forms of aqueous ammoniatedshellac dispersions are also commercially available, for exampleCertiseal® FC 300A film coat product, manufactured by Mantrose Haeuser,a subsidiary of RPM Corporation. Esterification of the shellac is alsolimited in these systems as shellac forms a salt with the ammonia orprotonated amino acid.

In another approach an enteric coating formulation in the form of aspray solution or suspension is used. This system may comprise shellacin aqueous salt form and sodium alginate, preferably in equalconcentrations. An aqueous solution of an alkali salt of shellac isprepared by first dissolving the shellac in 55° C. hot water, thenadding 10% ammonium hydrogen carbonate and heating to 60° C. andstirring for 30 minutes. Separately, a sodium alginate solution isprepared and the two solutions are then blended together. The system,when coated onto a dosage form rapidly disintegrates in simulatedintestinal fluid (pH 6.8).

The above approaches describe enteric coatings composed of food approvedingredients, which are either pH sensitive or more time dependent interms of their delayed release mechanism. However, all these systemsrequire multiple, time consuming preparation steps, often requiring twoseparate solutions to be made with additional dilution requirements andwhich increases the potential for error. Alternately, the systemsrequire the use of pre-made dispersions of EC or shellac, which thenrequire further dilution and blending steps thereby adding cost,complexity and/or time to the manufacturing process.

In the case of pre-made aqueous dispersions, a further cost is incurreddue to the need to store and ship dispersions which contain the addedbulk of water. Additionally, these pre-made aqueous dispersions requireadditional precautions to be taken to control microbial contaminationand to minimize any physical and/or chemical instability of thedispersion.

In some embodiments a formulation in powder form useful for producing asprayable dispersion for enteric coating is used. The powder formulationcomprising a food grade shellac, a non-ammonium alkali salt, andoptionally a water-miscible polymer. The powder formulation whendispersed in water is capable of producing a sprayable dispersion forenteric coating. This coating at 15% solids in water has a viscosity ofbelow 500 cps at about 25° C. when measured with a Brookfield LTVviscometer with a #2 spindle at 100 rpm.

In some embodiments the enteric coating comprises food grade shellac,optionally blended with other food grade ingredients. The coating isproduced starting from a powder form dispersed in water. In addition toshellac, the coating may comprise a non-ammonium alkali salt, which maybe selected from sodium bicarbonate, sodium carbonate, potassiumcarbonate, potassium bicarbonate, calcium hydroxide, calcium bicarbonateand calcium carbonate, and optionally a water-miscible polymer. Thewater-miscible polymer may be a polymer which is “food grade”,dissolvable or dispersible in water, with no discernable phaseseparation from the aqueous phase. The water-miscible polymers that maybe used include alginate salt, alginic acid, proteins (e.g. wheat,soybean or corn), methylcellulose (MC), hydroxypropylcellulose (HPC),hydroxypropylmethylcellulose (HPMC), carboxymethyl cellulose (CMC),pectin, carrageenan, guar gum, locust bean gum, xanthan gum, gellan gum,arabic gum, etc. In some embodiments the water-miscible polymer isselected from anionic polymers such as sodium carboxymethyl cellulose(CMC), sodium alginate or pectin. The coating may optionally compriseone or more plasticizers chosen from glycerine, mineral oil, triacetin,polyethylene glycol, glyceryl monostearate, acetylated monoglyceride,glyceryl tricaprylate/caprate and polysorbate. Optionally, the coatingmay further comprise pigments, and/or detackifiers such as titaniumdioxide, talc, iron oxide glyceryl monostearate. Additional componentssuch as natural colors, various carbohydrate derivatives such ashypromellose, hydroxypropyl cellulose, carboxymethyl starch, carageenanand xanthan may also be present.

The non-ammonium alkali salt used in the enteric coating composition maybe any food grade, nonvolatile, water soluble inorganic or organic saltspecies. The non-ammonium alkali salt may be selected from sodium,potassium, calcium, magnesium, aluminum salts. In some embodiments thenon-ammonium alkali salt comprises sodium bicarbonate. The amount ofnon-ammonium alkali salt of use in the enteric coating of is typicallyin the range of from 1.5% to 15% by weight of the coating, such as from1.5% to 8% by weight of the coating.

If the coating comprises a plasticizer, the plasticizer may be selectedfrom glycerine, propylene glycol, mineral oil, triacetin, polyethyleneglycol, acetylated monoglyceride, glyceryl monostearate, glyceryltricaprylate/caprate, polysorbate and oleic acid. Various edible oilsmay also serve as the plasticizers. The plasticizer may also be amedium-chain triglyceride which is a medium-chain (6 to 12 carbons)fatty acid ester of glycerol. If glycerine is the plasticizer, then itmay be used in an amount in the range of from about 1% to about 10% byweight, such as from 2% to 6% by weight. If mineral oil is theplasticizer, then it may be used in an amount in the range of from 3% to9%, such as from 5% to 7% by weight. If glyceryl monostearate is theplasticizer, then it may be used in an amount in the range of from 3% to25%, such as about 5% to about 20% by weight. If polysorbate 80 is theplasticizer, then it may be used in an amount in the range of from 0.5%to 12%, such as from 2% to 10% by weight. If acetylated monoglyceride isthe plasticizer, then it may be used in an amount in the range of from2% to 12%, such as from 4% to 10% by weight.

In some embodiments the enteric coating protects the active agent untilit reaches the distal ileum so that the active agent is then released inthe distal ileum or the distal ileum and the large intestine, resultingin addition of oxygen to the large intestine in an amount sufficient tomodify the microbiome of the large intestine. In some embodiments theenteric coating protects the active agent until it reaches the largeintestine so that the active agent is then released in the largeintestine, resulting in addition of oxygen to the large intestine in anamount sufficient to modify the microbiome of the large intestine. Insome embodiments the enteric coating protects the active agent until itreaches the distal ileum and/or large intestine and so that it is thenreleased over a period of at lease three hours, at least six hours, orat least twelve hours to thereby modify the microbiome of the largeintestine for a period of time.

In some embodiments of the particles, the weight ratio of delayedrelease coating to core is from 5% to 50%. In some embodiments of theparticles the weight ratio of delayed release coating to core is from20% to 30%. In some embodiments of the particles the weight ratio ofdelayed release coating to core is from 30% to 40%. In some embodimentsof the particles the weight ratio of delayed release coating to core isfrom 40% to 50%.

In some embodiments the delayed release coating prevents release untilthe active agent reaches the small intestine. In some embodiments thedelayed release coating prevents release until the active agent reachesthe ileum. In some embodiments the delayed release coating preventsrelease until the active agent reaches the distal ileum. In someembodiments the delayed release coating prevents release until theactive agent reaches the colon.

In some embodiments the delayed release coating is an enteric coatingthat is protective at pH below 5.5. In some embodiments the entericcoating is protective at pH below 6.5. In some embodiments the entericcoating is protective at pH below 7.0. In some embodiments the entericis protective at pH below 7.4. In some embodiments the delayed releasecoating is Eudagrit FS30D.

In some embodiments the particles comprise a core, a delayed releasecoating encapsulating the core, and a subcoat layer that encases thecore and is disposed between the core and the delayed release coating.The subcoat layer may serve to protect the core during manufacturing andin particular application of the delayed release coating. Alternatively,or in addition, the subcoat layer may serve as a release rate modifier.Thus, the subcoat layer may be chosen from any release rate modifierknown in the art, including any release rate modifier disclosed herein.In alternative embodiments the subcoat layer does not have release ratemodification properties. Nonlimiting examples of subcoat layers includeHPMC and guar gum.

In some embodiments the particles further comprise a subcoat layer thatencases the core and is disposed between the core and the entericcoating. In some embodiments the subcoat layer comprises HPMC.

In some embodiments the plurality of particles is encased in a capsuleor sachet. The capsule or sachet is generally chosen to deliver theplurality of particles to the stomach. In most embodiments the capsuleor sachet is of a type that immediately releases the contents into thestomach.

In some embodiments of the pharmaceutical preparations the plurality ofparticles is suspended in liquid for oral administration.

In some embodiments of the pharmaceutical preparations the plurality ofparticles is compressed with an excipient to form a tablet for oraladministration.

In some embodiments of the pharmaceutical preparations the plurality ofparticles is encased in a capsule or suspended in liquid for rectaladministration as a suppository.

In some embodiments the pharmaceutical preparation comprises no morethan one active agent. In some embodiments the pharmaceuticalpreparation comprises at least two active agents. In some embodimentsthe pharmaceutical preparation comprises at least three active agents.In embodiments comprising more than one active agent, each of theplurality or particles in the preparation may comprise each of theactive agents, or the preparation may comprise a plurality of types ofparticles, each type comprising a different active agent and/or set ofactive agents.

In some embodiments the pharmaceutical preparation comprises an activeagent increases pH. In some embodiments the pharmaceutical preparationcomprises an active agent comprises a bicarbonate group. In someembodiments pharmaceutical preparation comprises a bicarbonate salt,such as sodium bicarbonate, potassium bicarbonate, caesium bicarbonate,magnesium bicarbonate, calcium bicarbonate, ammonium bicarbonate, or amixture thereof.

In some embodiments the pharmaceutical preparation comprises an activeagent increases oxygen tension. In some embodiments active agentcomprises a peroxide functional group. In some embodiments the activeagent comprises hydrogen peroxide. In some embodiments the active agentis hydrogen peroxide. In some embodiments the active agent comprises anorganic peroxide. In some embodiments the active agent is an organicperoxide selected from acetyl acetone peroxide, acetyl benzoyl peroxide,ascaridole, benzoyl peroxide, di-(1-naphthoyl)peroxide, diacetylperoxide, ethyl hydroperoxide, ergesterol peroxide, iodoxy compounds,methyl isobutyl ketone peroxide. In some embodiments the pharmaceuticalpreparation comprises an active agent that is an inorganic peroxide. Insome embodiments the pharmaceutical preparation comprises an activeagent that is carbamide peroxide. In some embodiments the pharmaceuticalpreparation comprises an active agent that is sodium percarbonate.

In some embodiments the pharmaceutical preparation comprises an activeagent that increases production of oxygen from stores in the gut. Anexample is catalase, a common enzyme found in nearly all livingorganisms exposed to oxygen. Accordingly, In some embodiments thepharmaceutical preparation comprises an active agent that is a catalase.In some embodiments the catalase is a human catalase. In someembodiments the catalase is a non-human catalase. In some embodimentsthe catalase is a mammalian catalase. In some embodiments the catalaseis a non-mammalian catalase. In some embodiments the catalase is used asthe only agent that increases oxygen tension. In such embodimentscatalase increases oxygen tension by increasing the rate ofdecomposition of endogenous hydrogen peroxide to water and oxygen. Inother embodiments catalase and an active agent that comprises hydrogenperoxide are used together. In such embodiments the catalase increasesthe rate of production of oxygen from the hydrogen peroxide activeagent. In such embodiments the catalase may also act at least in part byincreasing production of oxygen from endogenous sources.

In some embodiments the pharmaceutical preparation comprises a firstactive agent that increases pH and a second active agent that increasesoxygen tension. In some embodiments the pharmaceutical preparationcomprises a first active agent that increases pH, a second active agentthat increases oxygen tension, and a third active agent that increasesproduction of oxygen from endogenous sources. In some embodiments thepharmaceutical preparation comprises a first active agent that increasespH, a second active agent that increases oxygen tension and is amolecule comprising a peroxide functional group, and a third activeagent that increases the rate of decomposition of the peroxidefunctional group to water and oxygen.

In some embodiments the pharmaceutical preparation comprises an activeagent that comprises a peroxide functional group. In some embodimentsthe active agent that comprises a peroxide functional group is hydrogenperoxide. In some embodiments the active agent that comprises a peroxidefunctional group is carbamide peroxide. In some embodiments the activeagent that comprises a peroxide functional group is sodium percarbonate.In some embodiments the pharmaceutical preparation further comprises aperoxide catalyst.

In some embodiments the pharmaceutical preparation comprises an activeagent that comprises a bicarbonate group. In some embodiments the activeagent that comprises a bicarbonate group is a bicarbonater salt. In someembodiments the active agent that comprises a bicarbonate group issodium bicarbonate.

In some embodiments the pharmaceutical preparation comprises a firstactive agent that comprises a peroxide functional group and a secondactive agent that comprises a bicarbonate group. In some embodiments thefirst active agent that comprises a peroxide functional group ishydrogen peroxide. In some embodiments the first active agent thatcomprises a peroxide functional group is carbamide peroxide. In someembodiments the first active agent that comprises a peroxide functionalgroup is sodium percarbonate. In some embodiments the second activeagent that comprises a bicarbonate group is a bicarbonater salt. In someembodiments the second active agent that comprises a bicarbonate groupis sodium bicarbonate. In some embodiments the pharmaceuticalpreparation further comprises a peroxide catalyst.

In some embodiments the pharmaceutical preparation is designed to targetrelease of the at least one active agent to the small intestine. In someembodiments the pharmaceutical preparation is designed to target releaseof the at least one active agent to the proximal small intestine. Insome embodiments the pharmaceutical preparation is designed to targetrelease of the at least one active agent to the distal small intestine.In some embodiments the pharmaceutical preparation is designed to targetrelease of the at least one active agent to the ileum. In someembodiments the pharmaceutical preparation is designed to target releaseof the at least one active agent to the distal ileum.

In some embodiments the pharmaceutical preparation is designed to targetrelease of the at least one active agent to the colon. In someembodiments the pharmaceutical preparation is formulated so that the atleast one active agent is released in the distal small intestine. Insome embodiments the pharmaceutical preparation is formulated so thatthe at least one active agent is released in the distal small intestineand the colon. In some embodiments the pharmaceutical preparation isformulated so that the at least one active agent is released in theileum. In some embodiments the pharmaceutical preparation is formulatedso that the at least one active agent is released in the ileum and thecolon. In some embodiments the pharmaceutical preparation is formulatedso that the at least one active agent is released in the distal ileum.In some embodiments the pharmaceutical preparation is formulated so thatthe at least one active agent is released in the distal ileum and thecolon. In some embodiments the pharmaceutical preparation is formulatedso that the at least one active agent is released in the jejunum. Insome embodiments the pharmaceutical preparation is formulated so thatthe at least one active agent is released in the colon.

In some embodiments the core does not comprise an enzyme. In someembodiments the core does not comprise an enzyme that oxidizes ethanolto acetate.

E. Methods of Weight Management

As demonstrated in the examples, the inventor has discovered thatadministering a pharmaceutical composition of the invention to a subjecthas a beneficial effect on the colon microbiome of the subject. Byincreasing the oxygen tension and/or redox potential and/or pH the colonenvironment is modified such that (1) the relative abundance ofbacterial types that promote at least one condition selected from ametabolic disease or disorder (such as obesity), metabolic syndrome, andcardiovascular disease is reduced; and/or (2) the relative abundance ofbacterial types that ameliorate at least one condition selected from ametabolic disease or disorder (such as obesity), metabolic syndrome, andcardiovascular disease is increased. It has also been discovered thatincreasing the oxygen tension and/or redox potential and/or pH in thecolon environment of a subject acts to manage the weight of the subject.Without wishing to be bound by theory, it appears that microbiomemodifications caused by increasing the oxygen tension and/or redoxpotential and/or pH of a subject's colon in turn manage the weight ofthe subject. This dramatic result is surprising and demonstratesunexpected benefits of the invention.

Accordingly, methods of weight management in a subject are alsoprovided. In some embodiments the methods comprise administering aneffective amount of a pharmaceutical composition of the invention to thesubject to thereby manage the weight of the subject. In some embodimentsthe weight management comprises at least one of weight loss, maintenanceof weight, controlling weight gain, body mass index (BMI) reduction,maintenance of BMI, and controlling BMI gain. In some embodiments thesubject is overweight or obese. In some embodiments the subject has atleast one weight-related condition. In some embodiments the at least oneweight-related condition is selected from a metabolic disease ordisorder, metabolic syndrome, and cardiovascular disease. In someembodiments the at least one weight-related condition is selected fromhypertension, dyslipidemia, and type 2 diabetes. In some embodiments atleast one symptom of the at least one weight-related condition isameliorated.

In some embodiments administering an effective amount of apharmaceutical composition of the invention to the subject increases therelative abundance of at least one of aerobic bacteria and facultativelyanaerobic bacteria in the microbiota of the colon of the subject. Insome embodiments administering the effective amount to the smallintestine and/or large intestine of the subject of at least one agentthat increases oxygen tension and/or redox potential and/or pH in thecolon of the subject increases the relative abundance of bacteria in atleast one phylum selected from Proteobacteria, Bacteriodetes, andVerrucomicrobia is increased in the microbiota of the colon of thesubject. In some embodiments the relative abundance of bacteria in thephylum Proteobacteria is increased in the microbiota of the colon of thesubject. In some embodiments administering an effective amount of apharmaceutical composition of the invention to the subject decreases therelative abundance of bacteria in the Firmicutes phylum in themicrobiota of the colon of the subject. In some embodimentsadministering an effective amount of a pharmaceutical composition of theinvention to the subject of at least one agent that increases oxygentension and/or redox potential and/or pH in the colon of the subjectdecreases the relative abundance of bacteria in the Firmicutes phylum inthe microbiota of the colon of the subject.

In some embodiments the at least one agent increases oxygen tension andis a peroxide. In some embodiments the at least one agent increases pHand is selected from bicarbonate salts, carbonate salts, bases, andbuffers. In some embodiments the at least one agent is a peroxidecatalyst. In some embodiments the at least one agent is selected fromcarbamide peroxide, sodium percarbonate, sodium bicarbonate, andcatalase. In some embodiments the method comprises administering aperoxide and catalase to the subject. In some embodiments the peroxideis selected from carbamide peroxide, sodium percarbonate. In someembodiments the method further comprises administering sodiumbicarbonate to the subject.

In some embodiments pharmaceutical composition of the invention isadministered to the subject without administering an enzyme to thesubject. In some embodiments of the methods the effective amount of apharmaceutical composition of the invention is administered to thesubject without administering an enzyme that oxidizes ethanol to acetateto the subject.

In general the pharmaceutical composition of the invention isadministered over a dosing period. A dosing period is a span of timeduring which the active agent is administered to a subject at a regulardosing interval, such as every four hours, every eight hours, everytwelve hours, or once a day. The dosing period may comprise one week,two weeks, one month, three months, six months, nine months, or oneyear. In some embodiments the active agent is administered at least oncea day for the dosing period. In some embodiments the active agent isadministered at least once every other day for the dosing period. Insome embodiments the active agent is administered at least two times aweek for the dosing period. In some embodiments the active agent isadministered at least three times a week for the dosing period. In someembodiments the active agent is administered at least four times a weekfor the dosing period. In some embodiments the active agent isadministered at least five times a week for the dosing period.

In some embodiments the dosing period is at least 3 days, at least 5days, at least 1 week, at least 2 weeks, at least 4 weeks, at least 2months, at least 3 months, at least 6 months, or at least one year. Insome embodiments the dosing period is 3 days, 5 days, 1 week, 2 weeks, 4weeks, 2 months, 3 months, 6 months, or one year. In some embodimentsthe dosing period is from 3 to 10 days, from 1 to 2 weeks, from 2 to 4weeks, from 1 to 2 months, from 3 to 6 months, or from 6 to 12 months.

In some embodiments the pharmaceutical composition of the invention isadministered for a dosing period and the weight of the subject does notincrease during the dosing period. In some embodiments thepharmaceutical composition of the invention is administered for a dosingperiod and the weight of the subject decreases during the dosing period.In some embodiments the weight of the subject decreases by at least 1%,at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, atleast 7%, at least 8%, at least 9%, or at least 10% during the dosingperiod.

In some embodiments the pharmaceutical composition of the invention isadministered for a dosing period and the BMI of the subject does notincrease during the dosing period. In some embodiments thepharmaceutical composition of the invention is administered for a dosingperiod and the BMI of the subject decreases during the dosing period. Insome embodiments the BMI of the subject decreases by at least 1%, atleast 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least7%, at least 8%, at least 9%, or at least 10% during the dosing period.

In some embodiments both the weight of the subject and the BMI of thesubject do not increase during the dosing period. In some embodimentsboth the weight of the subject and the BMI of the subject decreaseduring the dosing period. In some embodiments the weight of the subjectand the BMI of the subject independently each decrease by at least 1%,at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, atleast 7%, at least 8%, at least 9%, or at least 10% during the dosingperiod.

In some embodiments in which the subject has at least one weight-relatedcondition the at least one weight-related condition is selected from ametabolic disease or disorder, metabolic syndrome, and cardiovasculardisease.

In some embodiments the at least one weight-related condition ismetabolic syndrome. In some embodiments the metabolic syndrome ischaracterized by at least one feature selected from insulin resistance,central obesity, glucose intolerance, dyslipidemia with elevatedtriglycerides, low HDL-cholesterol, microalbuminuria, predominance ofsmall dense LDL-cholesterol particles, hypertension, endothelialdysfunction, oxidative stress, and inflammation. In some embodiments themetabolic syndrome is characterized by at least two features selectedfrom insulin resistance, central obesity, glucose intolerance,dyslipidemia with elevated triglycerides, low HDL-cholesterol,microalbuminuria, predominance of small dense LDL-cholesterol particles,hypertension, endothelial dysfunction, oxidative stress, andinflammation. In some embodiments the metabolic syndrome ischaracterized by at least three features selected from insulinresistance, central obesity, glucose intolerance, dyslipidemia withelevated triglycerides, low HDL-cholesterol, microalbuminuria,predominance of small dense LDL-cholesterol particles, hypertension,endothelial dysfunction, oxidative stress, and inflammation. In someembodiments the metabolic syndrome is characterized by at least fourfeatures selected from insulin resistance, central obesity, glucoseintolerance, dyslipidemia with elevated triglycerides, lowHDL-cholesterol, microalbuminuria, predominance of small denseLDL-cholesterol particles, hypertension, endothelial dysfunction,oxidative stress, and inflammation. In some embodiments the metabolicsyndrome is characterized by at least five features selected frominsulin resistance, central obesity, glucose intolerance, dyslipidemiawith elevated triglycerides, low HDL-cholesterol, microalbuminuria,predominance of small dense LDL-cholesterol particles, hypertension,endothelial dysfunction, oxidative stress, and inflammation.

In some embodiments the subject is pre-obese, obese, or morbidly obese.In some embodiments weight management of the pre-obese, obese, ormorbidly obese subject results in at least one of inducing weight lossin a pre-obese, obese, or morbidly obese subject; reducing BMI in apre-obese, obese, or morbidly obese subject; reducing food intake in apre-obese, obese, or morbidly obese subject; improving glucosehomeostasis in a pre-obese, obese, or morbidly obese subject; preventingweight gain and preventing an increase in BMI in a normal, pre-obese,obese, or morbidly obese subject.

In certain embodiments, the pharmaceutical composition of the inventionis administered to a subject suffering from obesity (e.g., a pro-obese,obese, or morbidly obese patient), an obesity-related disease ordisorder, diabetes, insulin-resistance syndrome, lypodystrpohy,nonalcoholic steatohepatitis, a cardiovascular disease, polycystic ovarysyndrome, or a metabolic syndrome.

In some embodiments the at least one condition is cardiovasculardisease.

In some embodiments the at least one condition is type-II diabetes.

Administering an effective amount to the small intestine and/or largeintestine of the subject of at least one agent that increases oxygentension and/or redox potential and/or pH may be achieved using anysuitable method. Generally, it will be preferable to provide the atleast one active agent in the form of an oral dosage form, including forexample any of the oral dosage forms disclosed herein. The oral dosageform is provided orally to the subject and then passes through thestomach to provide the at least one active agent to the small intestineand/or large intestine of the subject. However, alternative approachesmay also be used. For example, a feeding tube may be placed to provide acomposition comprising the at least one active agent directly to thesmall intestine or the large intestine. If an oral dosage form is used,any oral dosage form of this disclosure may be used, or any othersuitable oral dosage form known in the art.

In some embodiments the methods comprise administering no more than oneactive agent. In some embodiments the methods comprise administering atleast two active agents. In some embodiments the methods compriseadministering at least three active agents. In some embodiments of themethods a single oral dosage form is administered, which may comprise aplurality of active agents. In other embodiments a plurality of oraldosage forms is administered, which may each comprise a different activeagent or different combination of active agents.

In some embodiments the methods comprise administering an active agentthat increases pH. In some embodiments the methods compriseadministering an active agent that comprises a bicarbonate group and/ora carbonate group. In some embodiments the methods compriseadministering an active agent that comprises a bicarbonate salt, such aswithout limitation sodium bicarbonate, potassium bicarbonate, caesiumbicarbonate, magnesium bicarbonate, calcium bicarbonate, ammoniumbicarbonate, or a mixture thereof. In some embodiments the methodscomprise administering an active agent that comprises a carbonate salt.In some embodiments the methods comprise administering an active agentthat comprises a base. In some embodiments the methods compriseadministering an active agent that comprises a buffer.

In some embodiments the methods comprise administering an active agentthat increases oxygen tension. In some embodiments the methods compriseadministering an active agent that comprises a peroxide functionalgroup. In some embodiments the methods comprise administering an activeagent that comprises hydrogen peroxide. In some embodiments the methodscomprise administering an active agent that is hydrogen peroxide. Insome embodiments at least one agent is hydrogen peroxide. In someembodiments the at least one peroxide is an organic peroxide. In someembodiments the at least one peroxide is an organic peroxide selectedfrom acetyl acetone peroxide, acetyl benzoyl peroxide, ascaridole,benzoyl peroxide, di-(1-naphthoyl)peroxide, diacetyl peroxide, ethylhydroperoxide, ergesterol peroxide, iodoxy compounds, methyl isobutylketone peroxide. In some embodiments the at least one agent is aninorganic peroxide. In some embodiments the at least one agent is aninorganic peroxide selected from Ammonium persulfate, Calcium peroxide,Magnesium peroxide, Potassium persulfate, Sodium perborate, and Sodiumpercarbonate. In some embodiments the at least one agent is carbamideperoxide. In some embodiments the at least one agent is Sodiumpercarbonate. In some embodiments the methods comprise administering anactive agent that is carbamide peroxide. In some embodiments the methodscomprise administering an active agent that is Sodium percarbonate.

In some embodiments the methods comprise administering an active agentthat increases production of oxygen from stores in the gut and/or addedstores. In some embodiments the agent that increases production ofoxygen from stores in the gut and/or added stores is an organic peroxidecatalyst or an inorganic peroxide catalyst. An example is catalase, acommon enzyme found in nearly all living organisms exposed to oxygen.Accordingly, In some embodiments the methods comprise administering anactive agent that is a catalase. In some embodiments the catalase is ahuman catalase. In some embodiments the catalase is a non-humancatalase. In some embodiments the catalase is a mammalian catalase. Insome embodiments the catalase is a non-mammalian catalase. In someembodiments the catalase is used as the only agent that increases oxygentension. In such embodiments catalase increases oxygen tension byincreasing the rate of decomposition of endogenous hydrogen peroxide towater and oxygen. In other embodiments of the methods, catalase and anactive agent that comprises hydrogen peroxide are used together. In suchembodiments the catalase increases the rate of production of oxygen fromthe hydrogen peroxide active agent. In such embodiments the catalase mayalso act at least in part by increasing production of oxygen fromendogenous sources.

In some embodiments the methods comprise administering a first activeagent that increases pH and a second active agent that increases oxygentension. In some embodiments the methods comprise administering a firstactive agent that increases pH, a second active agent that increasesoxygen tension, and a third active agent that increases production ofoxygen from endogenous sources. In some embodiments the methods compriseadministering a first active agent that increases pH, a second activeagent that increases oxygen tension and is a molecule comprising aperoxide functional group, and a third active agent that increases therate of decomposition of the peroxide functional group to water andoxygen.

In some embodiments the methods comprise administering an active agentthat comprises a peroxide functional group. In some embodiments theactive agent that comprises a peroxide functional group is hydrogenperoxide. In some embodiments the active agent that comprises a peroxidefunctional group is carbamide peroxide. In some embodiments thepharmaceutical preparation further comprises a peroxide catalyst.

In some embodiments the methods comprise administering an active agentthat comprises a carbonate group or a bicarbonate group. In someembodiments the active agent that comprises a bicarbonate group is acarbonate salt or a bicarbonate salt. In some embodiments the activeagent that comprises a bicarbonate group is sodium bicarbonate. In someembodiments the active agent that comprises a carbonate group is sodiumcarbonate.

In some embodiments the methods comprise administering a first activeagent that comprises a peroxide functional group and a second activeagent that comprises a bicarbonate group. In some embodiments the firstactive agent that comprises a peroxide functional group is hydrogenperoxide. In some embodiments the first active agent that comprises aperoxide functional group is carbamide peroxide. In some embodiments thesecond active agent that comprises a bicarbonate group is a bicarbonatersalt. In some embodiments the second active agent that comprises abicarbonate group is sodium bicarbonate. In some embodiments thepharmaceutical preparation further comprises a peroxide catalyst.

The “effective amount” of the active agent(s) that is adminstered may bedetermined experimentally using methods that are standard in the art.For example, the methods described in the examples may be employed inmice and/or humans to identify an active agent and/or to perform adose-ranging study to define the range of amounts of the active agent(s)that provide a desired benefit and/or the lowest dose that provides adesired benefit.

For agents that increase oxygen tension a dose range of 0.1 ng/kg to 1g/kg may, for example, be tested in the mouse model to identify atherapeutically effective amount. For candidate agents that increase pHa dose range of 0.1 microgram/kg to 10 g/kg may, for example, be testedin the mouse model to identify a therapeutically effective amount.

Carbamide peroxide is used at a dose of from 1 ng/kg to 100 mg/kg perday, such as from 1 ng/kg to 10 ng/kg, from 10 ng/kg to 100 ng/kg, from1 microgram/kg to 10 micrograms/kg, from 10 microgram/kg to 100micrograms/kg, from 100 microgram/kg to 1 mg/kg, from 1 mg/kg to 10mg/kg, or from 10 mg/kg to 100 mg/kg, or at a dose that is bounded byany combination of two of these recited endpoints or at a dose of atleast one of these endpoints. The dose is administered 1, 2, 3, 4, 5, 6,7, 8, 9, or 10 times per day. In a preferred embodiment the dose isadministered 3 times per day.

In some embodiments carbamide peroxide is used at a dose of at least 25mg three times per day. In some embodiments carbamide peroxide is usedat a dose of at least 50 mg three times per day. In some embodimentscarbamide peroxide is used at a dose of at least 75 mg three times perday. In some embodiments carbamide peroxide is used at a dose of atleast 100 mg three times per day.

Sodium percarbonate is used at a dose of from 1 ng/kg to 100 mg/kg, suchas from 1 ng/kg to 10 ng/kg, from 10 ng/kg to 100 ng/kg, from 1microgram/kg to 10 micrograms/kg, from 10 microgram/kg to 100micrograms/kg, from 100 microgram/kg to 1 mg/kg, from 1 mg/kg to 10mg/kg, or from 10 mg/kg to 100 mg/kg, or at a dose that is bounded byany combination of two of these recited endpoints or at a dose of atleast one of these endpoints. The dose is administered 1, 2, 3, 4, 5, 6,7, 8, 9, or 10 times per day. In a preferred embodiment the dose isadministered 3 times per day.

In some embodiments sodium percarbonate is used at a dose of at least 25mg three times per day. In some embodiments sodium percarbonate is usedat a dose of at least 50 mg three times per day. In some embodimentssodium percarbonate is used at a dose of at least 75 mg three times perday. In some embodiments sodium percarbonate is used at a dose of atleast 100 mg three times per day.

In some embodiments the active agent comprises hydrogen peroxide and theagent is administered to a subject at a dose of from 0.01 mg to 100 mgof hydrogen peroxide equivalent. In some embodiments the active agent isadministered at a dose of from 0.1 mg to 5 mg of hydrogen peroxideequivalent. In some embodiments the active agent is administered at adose of from 5 mg to 10 mg of hydrogen peroxide equivalent. In someembodiments the active agent is administered at a dose of from 10 mg to20 mg of hydrogen peroxide equivalent. In some embodiments the activeagent is administered at a dose of from 20 mg to 40 mg of hydrogenperoxide equivalent. In some embodiments the active agent isadministered at a dose of from 40 mg to 60 mg of hydrogen peroxideequivalent. In some embodiments the active agent is administered at adose of from 60 mg to 80 mg of hydrogen peroxide equivalent. In someembodiments the active agent is administered at a dose of from 80 mg to100 mg of hydrogen peroxide equivalent.

In some embodiments the active agent is administered at a dose of from100 mg to 1 g of hydrogen peroxide equivalent. In some embodiments theactive agent is administered at a dose of from 100 mg to 200 mg ofhydrogen peroxide equivalent. In some embodiments the active agent isadministered at a dose of from 200 mg to 400 mg of hydrogen peroxideequivalent.

Sodium bicarbonate is used at a dose of from 1 microgram/kg to 1 g/kg,such as at 1 microgram/kg to 10 micrograms, from 10 micrograms/kg to 100micrograms/kg, from 100 micrograms/kg to 1 mg/kg, from 1 mg/kg to 10mg/kg, from 10 mg/kg to 100 mg/kg, or from 100 mg/kg to 1 g, or at adose that is bounded by any combination of two of these recitedendpoints or at a dose of at least one of these endpoints.

Catalase is used at a dose of from 0.0001 to 10,000 Baker's units perkg, such as from 0.0001 to 0.001 Baker's units per kg, from 0.001 to0.01 Baker's units per kg, from 0.01 to 0.1 Baker's units per kg, from0.1 to 1 Baker's units per kg, from 1 to 10 Baker's units per kg, from10 to 100 Baker's units per kg, from 100 to 1,000 Baker's units per kg,or from 1,000 to 10,000 Baker's units per kg. The dose is administered1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per day. In a preferredembodiment the dose is administered 3 times per day.

In some embodiments catalase is used at a dose of at least 25 mg threetimes per day. In some embodiments catalase is used at a dose of atleast 50 mg three times per day. In some embodiments catalase is used ata dose of at least 75 mg three times per day. In some embodimentscatalase is used at a dose of at least 100 mg three times per day.

In some embodiments the active agent is administered on an emptystomach. In other embodiments the active agent is administered withfood. In some embodiments the active agent is administered with meals.

In some embodiments of the methods the microbiota profile of the colonof the subject is modulated as a result of the treatment. In someembodiments administering the effective amount of the at least one agentto the small intestine and/or large intestine of the subject modulatesthe microbiota profile of the large intestine of the subject.

In some embodiments the relative abundance of at least one of aerobicbacteria and facultatively anaerobic bacteria is increased in themicrobiota of the colon of the subject. In some embodiments the relativeabundance of bacteria in at least one phylum selected fromProteobacteria, Bacteriodetes, and Verrucomicrobia is increased in themicrobiota of the colon of the subject. In some embodiments the relativeabundance of bacteria in the Proteobacteria phylum is increased in themicrobiota of the colon of the subject. In some embodiments the relativeabundance of bacteria in the Firmicutes phylum is decreased in themicrobiota of the colon of the subject. In some embodiments the relativeabundance of bacteria in at least one phylum selected fromProteobacteria, Bacteriodetes, and Verrucomicrobia is increased in themicrobiota of the colon of the subject; and the relative abundance ofbacteria in the Firmicutes phylum is decreased in the microbiota of thecolon of the subject.

In some embodiments the relative abundance of at least one class ofbacteria selected from Gammaproteobacteria, Alphaproteobacteria andBacteriodetes is increased in the microbiota of the colon of thesubject. In some embodiments the relative abundance of at least oneclass of bacteria selected from Clostridia, Erysipelotrichia, andBacilli is decreased in the microbiota of the colon of the subject. Insome embodiments the relative abundance of at least one class ofbacteria selected from Gammaproteobacteria, Alphaproteobacteria andBacteriodetes is increased in the microbiota of the colon of thesubject; and the relative abundance of at least one class of bacteriaselected from Clostridia, Erysipelotrichia, and Bacilli is decreased inthe microbiota of the colon of the subject.

In some embodiments the relative abundance of at least one order ofbacteria selected from Enterobacteriales, Bacteriodales, andVerrucomicrobiales is increased in the microbiota of the colon of thesubject. In some embodiments the relative abundance of at least oneorder of bacteria selected from Clostridiales, Erysiopelotrichales, andLactobacillales is decreased in the microbiota of the colon of thesubject. In some embodiments the relative abundance of at least oneorder of bacteria selected from Enterobacteriales, Bacteriodales, andVerrucomicrobiales is increased in the microbiota of the colon of thesubject; and the relative abundance of at least one order of bacteriaselected from Clostridiales, Erysiopelotrichales, and Lactobacillales isdecreased in the microbiota of the colon of the subject.

In some embodiments the relative abundance of at least one family ofbacteria selected from Enterobacteriaceae and Bacteroidaceae isincreased in the microbiota of the colon of the subject. In someembodiments the relative abundance of at least one family of bacteriaselected from Erysiopelotrichaceae, Prevotellaceae, Coriobacteriaceae,and Alcaligenaceae is decreased in the microbiota of the colon of thesubject. In some embodiments the relative abundance of at least onefamily of bacteria selected from Enterobacteriaceae and Bacteroidaceaeis increased in the microbiota of the colon of the subject; and therelative abundance of at least one family of bacteria selected fromErysiopelotrichaceae, Prevotellaceae, Coriobacteriaceae, andAlcaligenaceae is decreased in the microbiota of the colon of thesubject.

In some embodiments the relative abundance of at least one genus ofbacteria selected from Escherichia, Bacteroides, Alistipes, andAkkermansia is increased in the microbiota of the colon of the subject.In some embodiments the relative abundance of at least one genus ofbacteria selected from Clostridium and Lactobacillus is decreased in themicrobiota of the colon of the subject. In some embodiments the relativeabundance of at least one genus of bacteria selected from Escherichia,Bacteroides, Alistipes, and Akkermansia is increased in the microbiotaof the colon of the subject; and the relative abundance of at least onegenus of bacteria selected from Clostridium and Lactobacillus isdecreased in the microbiota of the colon of the subject.

In some embodiments the relative abundance of Akkermansia muciniphila isincreased in the microbiota of the colon of the subject.

In some embodiments the relative abundance of at least one type ofbacteria selected from Gammaproteobacteria, Alphaproteobacteria,Enterobacteriaceae (e.g., Escherichia), Bacteriodetes, andVerrucomicrobia (e.g., Akkermansia) is increased in the microbiota ofthe colon of the subject. In some embodiments the relative abundance ofat least one type of bacteria selected from Firmicutes (Clostridia),Erysiopelotrichaceae is decreased in the microbiota of the colon of thesubject.

In some embodiments the relative abundance of at least one type ofbacteria selected from Gammaproteobacteria, Alphaproteobacteria,Enterobacteriaceae (e.g., Escherichia), Bacteriodetes, andVerrucomicrobia (e.g., Akkermansia) is increased in the microbiota ofthe colon of the subject; and the relative abundance of at least onetype of bacteria selected from Firmicutes (Clostridia),Erysiopelotrichaceae is decreased in the microbiota of the colon of thesubject.

In some embodiments the methods further comprise collecting a stoolsample from the subject and assaying the microbiota profile in the stoolsample to determine the relative abundance of at least one phylum ofbacteria selected from Proteobacteria, Bacteriodetes, Verrucomicrobia,and Firmicutes; and/or to determine the relative abundance of at leastone class of bacteria selected from Gammaproteobacteria,Alphaproteobacteria, Bacteriodetes, Clostridia, Erysipelotrichia, andBacilli; and/or to determine the relative abundance of at least oneorder of bacteria selected from Enterobacteriales, Bacteriodales,Verrucomicrobiales, Clostridiales, Erysiopelotrichales, andLactobacillales; and/or to determine the relative abundance of at leastone family of bacteria selected from Enterobacteriaceae, Bacteroidaceae,Erysiopelotrichaceae, Prevotellaceae, Coriobacteriaceae, andAlcaligenaceae; and/or to determine the relative abundance of at leastone genus of bacteria selected from Escherichia, Bacteroides, Alistipes,Akkermansia, Clostridium, and Lactobacillus; and/or to determine therelative abundance of Akkermansia muciniphila. In some embodiments thestool sample is collected at least one time point selected from beforeinitiation of administering at least one agent that increases oxygentension and/or redox potential and/or pH, during a course ofadministering at least one agent that increases oxygen tension and/orredox potential and/or pH, and after completion of administering acourse of at least one agent that increases oxygen tension and/or redoxpotential and/or pH.

In some embodiments in which the relative abundance of a type ofbacteria is increased in the colon of the subject the increase inrelative abundance is at least 25%, at least 50%, at least 75%, at least100% (i.e., one fold), at least 200% (i.e., two fold), at least 300%, atleast 400%, at least 500%, at least 600%, at least 700%, at least 800%,at least 900%, at least 1,000% (i.e., ten fold), or at least 10,000%(i.e., 100 fold). In some embodiments in which the relative abundance ofa type of bacteria is decreased in the colon of the subject the decreasein relative abundance is at least 10%, at least 25%, at least 50%, atleast 75%, at least 90% (i.e., ten fold), at least 95% (i.e., 20 fold),at least 99% (i.e., 100 fold), at least 99.9% (i.e., 1000 fold).

In some embodiments of the methods the at least one agent that increasesoxygen tension and/or redox potential and/or pH in the colon of asubject is administered to the subject without administering an enzymeto the subject. In some embodiments of the methods the at least oneagent that increases oxygen tension and/or redox potential and/or pH inthe colon of a subject is administered to the subject withoutadministering an enzyme that oxidizes ethanol to acetate to the subject.

EXAMPLES

The examples are provided to further define the disclosure without,however, limiting the disclosure to the specifics of these examples.

Example 1: Pharmaceutical Preparations

A multiparticulate formulation composed of 3 components was formed,namely a core particle (˜1 mm diameter) containing the API, a subcoatlayer to create a barrier between the functional (enteric) coating andthe API, and finally the functional coating as the outmost layer, whichis an enteric coating designed to target the release of API to thedistal small intestine/proximal colon region.

Core API particle: Sodium percarbonate (Provox C, OCI ChemicalCorporation).

Subcoat: HPMC layer.

Enteric coating: Eudagrit FS30D.

Sodium percarbonate particles were coated first with a subcoat layer ofHPMC. Next, the particles having the subcoat were coated with HPMC (5%weight gain), and then coated with the Eudagrit FS3OD (35% weight gain).

Methods

Sodium percarbonate particles were prepared by sieving using a 0.6 mmsieve under dry conditions.

The HPMC layer coating solution in ddH₂O was prepared as follows usingHypromellose USP PHARMACOAT 606 (LOT No.: 5118397) and Super Refined PEG400-LQ-(MH), SR40377 (LOT No.: 0000608682). Briefly, 30.0 g HPMC (7.5%w/w) and 3.0 g (0.75% w/w) PEG 400 were dissolved in 400 g water.

Enteric coating layer solution in ddH₂O was prepared using the followingmaterials: EUDRAGIT® FS 30 D (LOT No.: B15065004), Talc USP (LOT No.:2DK0194), H₂O: Milli-Q, Triethyl citrate USP-NF (LOT No.: 6P013354),Glyceryl Monostearate NF (LOT No.: 127309), and Polysorbate 80 NF (LOTNo.: 2FB0143).

Two enteric coating formulas were prepared and tested. Formula 1:EUDRAGIT® FS 30D-250 g, Talc—37.5 g, H₂O 275—g, total=562.5 g total.Formula 2: EUDRAGIT® FS 30D-298.9 g, Polysorbate 80 (33%)— 4.4 g,Triethyl citrate—4.5 g, GMS—3.6 g, H₂O—188.7 g=500 g total.

In each case the coating solution was passed through a 0.25 mm sieve andcontinuously stirred using a magnetic stirrer during the coating processto prevent the solids from settling.

The sodium percarbonate particles were coated with an HPMC layer using afluid bed having the following settings: Preheating temperature (°C.)—45, Atomizing pressure—1.5-2, Outlet air temperature—40-45, Dryingtemperature—42, Capacity of fan—8-9, Pump speed (rpm)—1.6-2.5. Sodiumpercarbonate particles were sieved using a 0.6 mm sieve before coating.Particles larger than 0.6 mm were collected. Particle weight gain aftersealing layer coating was ˜5.0%.

The sealed sodium percarbonate particles were coat sealed with aEUDRAGIT® FS 30 D layer using a fluid bed having the following settings:Preheating temperature (° C.)— 40, Atomizing pressure—1.5-2, Outlet airtemperature—38-42, Drying temperature—40, Capacity of fan—8-9, Pumpspeed (rpm)—1.6-2.5.

Weight gain after enteric coating (Formula #1: FS30D+Talc): 6.8%, 12.7%,20.3%, 35.0%, 50.0%. Weight gain after enteric coating (Formula #2:FS30D+GMS): 24.9%. Size of coated particles was between 0.9 mm and 1.18mm. Average weight of each coated particle was 0.96±0.07 mg (N=10).35.0% and 50.0% weight gain particles are filled into size 3 HPMCcapsules (CAPSUGEL, Vcaps Plus Hypromellose Capsules) (200 mg/capsules)for further testing.

Results

A two-stage dissolution test was used. Stage 1: 0.1 M HCl (pH 1.2) for 2hours, Stage 2: 0.2 M PBS (pH 7.4) until dissolved, Paddle speed: 100rpm, Volume: 500 mL, Temperature: 37° C. Three individual vessels wereused for each sample (˜1 g particles/vessel). Enteric coatingdissolution was determined visually by presence or absence of gas bubblerelease from the particles.

Formula #1 was evaluated with two enteric coating levels: 6.8% weightgain and 12.7% weight gain. Neither remained stable in 0.1 M HCl for 2hours. But the 12.7% weight gain samples exhibited longer stability inacid.

The coating level was to ˜20.0% and the two enteric coating formulationswere compared. Stability in acid was significantly improved. Formula #1showed better acid stability than Formula #2. After changing medium to0.2 M PBS (pH 7.4), coated particles dissolved within 10 minutes.

Next enteric coating level using Formula #1 was increased to 35% and50%. Both coating levels protected API in 0.1 M HCl, with no dissolutionobserved. Although at 35% coating level, several particles started toflow on the surface of the dissolution medium after 1 hour. Afterchanging to 0.2 M PBS (pH 7.4), both groups of coated particles startedto dissolve within 15 minutes. Slight delay was observed in the 50%coating group. Completion of dissolution was noticed after 1 hour.

Due to the existence of Talc in formulation #1, dissolution mediumbecame turbid afterwards. The dissolution process proceeded as follows:particles first started to float on the medium surface, bubbles wereobserved on the surface of the coated particles as well as in themedium. Afterwards, more bubbles formed and dissolution medium startedturning turbid. Finally, all of the coated particles lost the rigidround shape and completely dissolved in PBS.

In vitro results for Formula #1 showed good enteric properties andsuccessful release at pH 7.4, indicating its suitability for use invivo.

In the following examples a formulation using 35% coating level versionof Formula #1 was used, as it showed good enteric properties.

Example 2: Methods of Weight Management

Methods

Three subjects participated in the study. The study followed an adaptivetrial design with dynamic dose adjustment. Subject 1 was a 57-year-oldfemale with a baseline body-mass index (BMI) of 23.6; Subject 2 was a56-year-old male with a baseline BMI of 27.9; and Subject 3 was a33-year-old male with a baseline BMI of 37.1. Dose range was 1-3mg/kg/day. Baseline anthropometric and clinical parameters for studysubjects are outlined in Table 1.

TABLE 1 Anthropometric and clinical parameters Subject #1 Subject #2Subject #3 Age, years 57 56 33 Gender, F/M F M M BMI, baseline, kg/m223.6 27.9 37.1 Weight, baseline, kg 60.5 90.4 131

Subjects were weight stable for at least three months prior to dosing.Subjects were not on any medications before (up to 1 month) or duringdosing. Table 2 describes the pill-dosing regimen used for each subject.Dose values indicate API amount.

TABLE 2 Dosing regimen Subject #1 Subject #2 Subject #3 Time period Day0-36 Day 0-10 Day 0-6 Dose frequency QD QD BID Dosing times AM AM AM/PMDose per administration, mg 100 100 100 Total daily dose, mg 100 100 200Total daily dose bw adj., mg/kg 1.67 1.11 1.53 Time period — Day 11-21Day 7-14 Dose frequency — BID QID Dosing times — AM/PM Every 6 hoursDose per administration, mg — 100 100 Total daily dose, mg — 200 400Total daily dose bw adj., mg/kg — 2.22 3.05 Time period — Day 22-34 Day15-36 Dose frequency — QD BID Dosing times — AM AM/PM Dose peradministration, mg — 200 200 Total daily dose, mg — 200 400 Total dailydose bw adj., mg/kg — 2.22 3.05 Total dosing period, days 36 34 36

Subjects were instructed to continue ad libitum diet during dosing (nochange in daily food intake, same as baseline) and they were instructednot to exercise in addition to their daily activities. Subjects werealso instructed not to consume any alcohol during dosing period.

Fasting blood glucose measurements: Fasting self-measured plasma glucose(SMPG) measurements were taken daily in the mornings. In FIG. 2, eachdaily data point for SMPG indicates the average of daily SMPGmeasurements over the previous 8 days. Subject #1 had 8 baseline datapoints for SMPG and subject #2 and #3 had only 1 baseline data point.Only subjects #1 and #2 consistently performed SMPG measurements duringdosing period.

Urine ketone measurements: Subjects performed daily urine ketonemeasurements using color indicator ketone strips (TRUEplus ketonestrips), which measure urine acetoacetate concentration (mg/dL). Eachmeasurement was recorded against the reference color scale andphotographed for independent scoring and future reference.

Serum metabolic marker measurements: Subjects #1 and #2 had venous blooddrawn at a medical center at dosing day 28 and sampled blood wasanalyzed for clinical parameters (liver, cholesterol, glucosemetabolism, thyroid hormones etc) as listed in Table 3.

TABLE 3 Metabolic data Subject #1 Subject #2 Subject #3 Body weight, %change −5.29% −4.76% −6.03% Absolute change, kg −3.2 −4.3 −7.9 Baseline,kg 60.5 90.4 131 Dose end, kg 57.3 86.1 123.1 BMI, absolute change,kg/m2 −1.3 −1.3 −2.2 Baseline, kg/m2 23.6 27.9 37.1 Dose end, kg/m2 22.426.6 34.8 % Excess weight loss, day 36 −46%    −29%    −16%    SMPG,8-day avg, % change −8.74% 2.40% — Absolute change, mg/dL −8.7 2.4 —Baseline, mg/dL 99.5 99.9 201 Dose end, mg/dL 90.8 102.3 — Fastinginsulin, day 28, uU/mL 3 9.7 — HbA1c (%), day 28 5.3 5.2 — HOMA-IR, day28 0.6 2.5 — ALT, day 28 13 20 — AST, day 28, U/L 14 18 — ALP, day 28 4544 — Bilirubin, day 28, mg/dL 0.48 0.75 — GGT, day 28, U/L 20 15 — T3Free, day 28, pg/mL 2.19 — — T4 Free, day 28, ng/dL 1.16 — — TSH, day28, mIU/L 1.76 — — Triglycerides, day 28, mg/dL 68 85 — HDL, day 28,mg/dL 89 56 — LDL, day 28, mg/dL 129 110 — Total cholesterol, day 28,mg/dL 232 183 —

Results

Bodyweight decreased substantially in all subjects after 5-6 weeks ofdosing (−5.36% mean change from baseline, −5.1 kg mean absolute change,16-46% excess weight loss, Table 3). Each subject's body weight overtime is plotted in FIG. 1. These results are on par with efficacy ofgastric bypass surgery (10-15% EWL/month, 4-5 kg/month) (1, 2). Thesubstantial weight loss of ˜15 kg (˜33 lbs; 11.5% loss from baseline)through the follow-up period in subject #3 (FIG. 4), who is a potentialcandidate for bariatric surgery, is particularly noteworthy. Based onself-reporting of all subjects, the pattern of weight loss was notattributable to changes in dietary habits (i.e., no change in absolutefood intake or food choice).

Food Intake and Hunger: Throughout dosing period, food intake or foodpreference reported by subjects did not change significantly and wassimilar to baseline (data not shown). Subjects reported loss of appetitein the first few days at the start of dosing but their appetite returnedto normal thereafter. Self reported hunger levels (average of last 2days, on a 1-10 visual scale (3)) decreased relative to baseline towardsthe end of dosing (baseline: 5.6 (0.7), end of dosing: 3.7 (0.4)).

Metabolic Parameters: All the metabolic data is summarized in Table 3.FIG. 2 shows self-measured plasma glucose (SMPG) (8-day average) andurine ketone results for subjects #1 and #2. FIG. 3 shows urine ketoneresults for subject #3 over dosing period. Subject #1 had a decrease inSMPG compared to baseline at end of dosing, whereas subject #2 had aslight increase. All subjects experienced substantial increase in ketonebody excretion in urine during dosing period indicating ketogenesis, fatmobilization and fat oxidation. Urine and serum ketones also increasedramatically in the early post-operative period (0-6 months) after RYGB(4, 5).

All subjects experienced significant increases in ketone body excretionin urine as measured by in-home urinalysis (TRUEplus™ ketone strips)during the dosing period indicating induction of ketogenesis (i.e.,physiological ketosis), fat mobilization and fat oxidation. Of note,urine and serum ketones also increase dramatically in the earlypost-operative period (0-6 months) after RYGB surgery. (14, 15) 16S rRNAsequencing of fecal samples was also performed in Subject #3 using amicrobiome sampling kit (OMNIgene-Gut, DNA Genotek). The results showedthat relative abundance of Proteobacteria increased to 6.14% (day 3 ofdoing) from 3.73% (baseline) in Subject #3.

The original test plan was simply to gather an initial understanding oftolerability and potential efficacy. Therefore, no baselines were takenfor other measures besides weight. However, the 5.4% reduction in weightfollowing 28 days allowed for a unique opportunity to assess some othermeasures in an antidotal manner. Two of the non-obese test subjectssubmitted themselves for the testing of liver markers (ALT, AST, ALP,GGT, Bilirubin) on dosing day 28. The results were within normal range,indicating favorable liver safety profile (Table 3). Fasting insulinlevels were 3 and 9.7 uU/mL for subjects #1 and #2 respectively.

LDL cholesterol was in the high range (>100 mg/dL) in both subjects,which could be due to active fat mass loss and fat mobilization intoserum (6, 7), which is also supported by constantly high urine ketonelevels independent of food intake (FIGS. 2,3). These results areconsistent with low fasting insulin levels measured at day 28 (Table 3)(8). In particular, subject #1 had extremely low insulin levels comparedto average female in the same BMI range (3 uU/mL vs 6-8 uU/mL) (9).After RYGB, insulin levels also rapidly fall and insulin resistanceresolves (10, 11).

Though no direction of change could be determined due to a lack ofbaseline measurements for these markers, the high range of the LDL-clevels may suggest that active fat mobilization caused a transientincrease in this marker. This is consistent with a phenomenon that hasbeen reported in non-obese subjects during weight loss. (16, 17) Thisstate of fat mobilization is also in line with constantly high urineketone levels independent of food intake observed in our test subjects.These results are also consistent with relatively low range fastinginsulin levels measured at day 28 in the study group and as previouslydemonstrated during weight loss. (18) Further to this hypothesis, afterRYGB, insulin levels also rapidly fall and insulin resistance resolves.(19, 20)

The most frequent AEs were constipation, dry mouth, metallic taste inmouth, burnt sensation on tongue, loss of appetite and increased thirst& water consumption. All AEs, except constipation and increased waterconsumption, resolved after the first 1-2 weeks of dosing. These AEs areconsistent with a prolonged ketosis state achieved during dosing, asfasting or low carb diet induced ketosis also produces similar sideeffects (12). In particular, metallic taste in mouth during ketosis isbelieved to be a result of a ketone body acetone being excreted into themouth during ketosis. There were no significant AE frequency or severitydifferences between dosing levels or subjects

Discussion

This proof-of-concept clinical trial tested the safety, tolerability,and weight-reducing effects of a novel oral formulation designed tomimic the mechanism of action of gastric bypass surgery. The formulationappeared to be safe over 5 weeks of dosing with minor side effects. Therate and extent of weight loss, and metabolic data reported in thisstudy should be contextualized. The minimum clinically significantweight loss after 1 year of therapy (behavioral modification and/ormedication) is often regarded as 3-5%, assuming risk factors such aswaist circumference, blood pressure, serum lipids and inflammatorymarkers also improve. In fact, the US FDA has set 5% total bodyweightloss at 1 year as a clinical efficacy hurdle for marketing approval ofdrugs intended to treat obesity. In the current study, our formulationresulted in profound bodyweight loss of approximately 5-6% (16-46% EWL)at 5 weeks. Remarkably, the observed weight loss was achieved with nosignificant change in food intake, suggesting increased basal energyexpenditure. These results are in line with the efficacy of bariatricsurgery. RYGB patients usually lose 4-5% of bodyweight (10-20% EWL) inthe first month on a very low calorie diet (300-600 kcal/day). A largefraction of this initial weight loss is actually due to the trauma ofthe surgery and includes significant lean mass loss (1, 2). We alsoobserved rapid appearance and sustained presence of ketone bodies inurine despite relatively constant food intake. This observation stronglysuggests dosing-induced drop in basal insulin levels and subsequent fatmobilization, lipolysis and fat oxidation (13).

Example 3: Methods of Weight Management

A rodent model of obesity was used to further evaluate the efficacy ofFormula 1. Sprague-Dawley male (12 weeks old) rats that were fed eithernormal chow or a HFD, and animals were treated daily by gavage witheither Formula 1 (pH 7 Formulation) or vehicle controls. There were 6animals in each of the four treatment groups (i.e., chow+vehicle,chow+FORMULA 1, HFD+vehicle, HFD+FORMULA 1). FORMULA 1 treatment dosewas 33 mg/kg, which is approximately ˜⅓ of theno-observable-adverse-effect-level (NOAEL; 81-115 mg/kg/day) for sodiumpercarbonate based on chronic oral hydrogen peroxide treatment in rat.(21) Animals treated with FORMULA 1 gained the same amount of weight asdid the chow+vehicle control group, while the HFD+vehicle group gainedsignificantly more weight than the others at 21 days (20% vs. 6-9%). Alarge variability in animals dosed with FORMULA 1 on HFD in terms ofefficacy was, however, noted. Half (n=3) of the animals showed strongefficacy while the other half (n=3) of the rats did not; therefore, thislatter subgroup is separated out into their own group (“fail group”) inthe results of this study presented in FIG. 5. Without wishing to bebound by any theory, it is presently believed that the lack of responsein this group is due to the unoptimized nature of the pH 7 Formulationfor rat physiology. Among other GI differences from human, rats have alower pH in the intestine (upper range ˜5-7) compared to human (upperrange ˜6.5-7.4) (22) and thus it is believed that the pH 7 Formulationused in this study failed to release API consistently in all the ratsdue to inter-rat variability at the upper normal range of intestinal pH(i.e., not all rats had a pH of 7 in their intestines). Hence, the starkbinary outcome data in the FORMULA 1 high fat dosing group may reflectthe variation of API delivery success, due to the specific physiologicalcharacteristics of the rat GI tract (pH, transit time, water contentetc.).

Variability in intestinal pH aside, the results of this study areconsistent with the high efficacy that the API displayed in humans.Similarly, RYGB surgery performed in rat models prevents weight gain inHFD fed animals, but has no effect on body weight in lean, chow fedanimals. It can be hypothesized that the stark nature of this binaryoutcome result in rats (high efficacy in half, no efficacy in the otherhalf) reflects a failure of the formulation (unoptimized coating for therat intestine), instead of a more graded result (i.e. normaldistribution), which would be expected from true physiological variationin response to the encapsulated API. It is also salient to point outthat even if the efficacy rates observed with Formula 1 in rats were totranslate exactly to humans in large trials, this invention would stillbecome an extremely successful commercial product, as almost allmedicines—including surgery (RYGB)—have large efficacy variation inhuman populations, and there are currently no effective weight lossagents that can achieve efficacy rates on par RYGB.

REFERENCES

-   1. A. Mor, L. Sharp, D. Portenier, R. Sudan, A. Torquati, Weight    loss at first postoperative visit predicts long-term outcome of    Roux-en-Y gastric bypass using Duke weight loss surgery chart, Surg    Obes Relat Dis 8, 556-60 (2012).-   2. C. Ciangura, J.-L. L. Bouillot, C. Lloret-Linares, C. Poitou, N.    Veyrie, A. Basdevant, J.-M. M. Oppert, Dynamics of change in total    and regional body composition after gastric bypass in obese    patients, Obesity (Silver Spring) 18, 760-5 (2010).-   3. A. Flint, A. Raben, J. E. Blundell, A. Astrup, Reproducibility,    power and validity of visual analogue scales in assessment of    appetite sensations in single test meal studies, Int. J. Obes.    Relat. Metab. Disord. 24, 38-48 (2000).-   4. N. Friedrich, K. Budde, T. Wolf, A. Jungnickel, A. Grotevendt, M.    Dressler, H. Völzke, M. Blüher, M. Nauck, T. Lohmann, H.    Wallaschofksi, Short-term changes of the urine metabolome after    bariatric surgery, OMICS 16, 612-20 (2012).-   5. E. Gralka, C. Luchinat, L. Tenori, B. Ernst, M. Thurnheer, B.    Schultes, Metabolomic fingerprint of severe obesity is dynamically    affected by bariatric surgery in a procedure-dependent manner,    Am. J. Clin. Nutr. 102, 1313-22 (2015).-   6. J. C. Swaner, W. E. Connor, Hypercholesterolemia of total    starvation: its mechanism via tissue mobilization of cholesterol,    Am. J. Physiol. 229, 365-9 (1975).-   7. L. Savendahl, L. E. Underwood, Fasting increases serum total    cholesterol, LDL cholesterol and apolipoprotein B in healthy,    nonobese humans, J. Nutr. 129, 2005-8 (1999).-   8. M. D. Jensen, M. Caruso, V. Heiling, J. M. Miles, Insulin    regulation of lipolysis in nondiabetic and IDDM subjects, Diabetes    38, 1595-601 (1989).-   9. L. P. Palaniappan, Heterogeneity in the Relationship Between    Ethnicity, BMI, and Fasting Insulin, DIABETES CARE 25 (2002).-   10. K. Wickremesekera, G. Miller, T. D. Naotunne, G. Knowles, R. S.    Stubbs, Loss of insulin resistance after Roux-en-Y gastric bypass    surgery: a time course study, Obes Surg 15, 474-81 (2005).-   11. A. P. Chambers, L. Jessen, K. K. Ryan, S. Sisley, H. E.    Wilson-Pérez, M. A. Stefater, S. G. Gaitonde, J. E. Sorrell, M.    Toure, J. Berger, D. A. D'Alessio, S. C. Woods, R. J. Seeley, D. A.    Sandoval, Weight-independent changes in blood glucose homeostasis    after gastric bypass or vertical sleeve gastrectomy in rats,    Gastroenterology 141, 950-8 (2011).-   12. T. C. Crowe, Safety of low-carbohydrate diets, Obes Rev 6,    235-45 (2005).-   13. D. W. Foster, J. D. McGarry, The regulation of ketogenesis, Ciba    Found. Symp. 87, 120-31 (1982).-   14. Friedrich N, Budde K, Wolf T, et al. Short-term changes of the    urine metabolome after bariatric surgery. OMICS 2012; 16:612-20.-   15. Gralka E, Luchinat C, Tenori L, Ernst B, Thurnheer M,    Schultes B. Metabolomic fingerprint of severe obesity is dynamically    affected by bariatric surgery in a procedure-dependent manner. Am J    Clin Nutr 2015; 102:1313-22.-   16. Swaner J C, Connor W E. Hypercholesterolemia of total    starvation: its mechanism via tissue mobilization of cholesterol. Am    J Physiol 1975; 229:365-9.-   17. Savendahl L, Underwood L E. Fasting increases serum total    cholesterol, LDL cholesterol and apolipoprotein B in healthy,    nonobese humans. J Nutr 1999; 129:2005-8.-   18. Jensen M D, Caruso M, Heiling V, Miles J M. Insulin regulation    of lipolysis in nondiabetic and IDDM subjects. Diabetes 1989;    38:1595-601.-   19. Wickremesekera K, Miller G, Naotunne T D, Knowles G, Stubbs R S.    Loss of insulin resistance after Roux-en-Y gastric bypass surgery: a    time course study. Obes Surg 2005; 15:474-81.-   20. Chambers A P, Jessen L, Ryan K K, et al. Weight-independent    changes in blood glucose homeostasis after gastric bypass or    vertical sleeve gastrectomy in rats. Gastroenterology 2011;    141:950-8.-   21. OECD. SODIUM PERCARBONATE. In: Organisation for Economic    Co-operation and Development (OECD).-   22. Peters S A. Physiologically-Based Pharmacokinetic (PBPK)    Modeling and Simulations: Principles, Methods, and Applications in    the Pharmaceutical Industry, First Edition: John Wiley & Sons, Inc.;    2012.

1-41. (canceled)
 42. A composition comprising a plurality of particles,each particle comprising: 1) a core comprising an oxygen-releasingagent, and 2) an enteric coating encasing the core.
 43. The compositionof claim 42, wherein the composition comprises a capsule or sachetencasing the plurality of particles.
 44. The composition of claim 43,wherein the oxygen-releasing agent is characterized in that it increasesthe oxygen tension of a solution of phosphate buffered saline.
 45. Thecomposition of claim 43, wherein the composition is characterized inthat it achieves release of oxygen when assayed in PBS buffer at pH 7.4after immersion at pH 1.2 for 2 hours.
 46. The composition of claim 44or 45, wherein enteric coating dissolves at a pH above 5.5, 5.6, 6.0,6.5, 6.8, 7.0, or 7.4.
 47. The composition of claim 44 or 45, whereinenteric coating dissolves at a pH between 6.8 and 7.2.
 48. Thecomposition of claim 44 or 45, wherein enteric coating dissolves at a pHabove 7.0.
 49. The composition of claim 44 or 45, wherein entericcoating comprises Eudragit FS 30D or Eudragit S100.
 50. The compositionof claim 44 or 45, wherein the enteric coating comprises methacrylicacid copolymers, polyvinyl acetate phthalate, cellulose acetatephthalate, hydroxypropyl methylcellulose phthalate, or hydroxypropylmethylcellulose acetylsuccinate.
 51. The composition of claim 44 or 45,wherein the enteric coating comprises a solid powder of an anioniccopolymer matrix based on methacrylic acid and methyl methacrylate. 52.The composition of claim 44 or 45, wherein the composition ischaracterized in that, when orally administered to a human subject, atleast 50% of the oxygen-releasing agent is released over a period of 4hours after contacting the small intestine.
 53. The composition of claim44 or 45, wherein the oxygen-releasing agent comprises a peroxide. 54.The composition of claim 53, wherein the peroxide is selected from thegroup consisting of: (i) hydrogen peroxide; (ii) an organic peroxideselected from acetyl acetone peroxide, acetyl benzoyl peroxide,ascaridole, benzoyl peroxide, di-(1-naphthoyl)peroxide, diacetylperoxide, ethyl hydroperoxide, ergesterol peroxide, and methyl isobutylketone peroxide; (iii) an inorganic peroxide selected from ammoniumpersulfate, calcium peroxide, magnesium peroxide, Potassium persulfate,and sodium percarbonate; and (iv) carbamide peroxide.
 55. Thecomposition of claim 53, wherein the peroxide is selected from carbamideperoxide and sodium percarbonate.
 56. The composition of claim 42,wherein the oxygen-releasing agent comprises sodium percarbonate. 57.The composition of claim 44 or 45, wherein the composition does notcomprise an enzyme that oxidizes ethanol to acetate.
 58. The compositionof claim 44 or 45, wherein the composition does not comprise an enzymethat catalyzes decomposition of hydrogen peroxide.
 59. The compositionof claim 44 or 45, wherein the composition does not comprise catalase.60. The composition of claim 44 or 45, wherein the cores of theparticles have a maximum dimension of from 0.1 to 2 mm.
 61. Thecomposition of claim 44 or 45, wherein the cores of the particles have amaximum dimension of from 0.2 to 1.0 mm.
 62. The composition of claim 44or 45, wherein the particles have a maximum dimension of 3.0 mm or less.63. The composition of claim 44 or 45, wherein the composition comprisesfrom 25 to 500 mg of the oxygen-releasing agent.
 64. The composition ofclaim 44 or 45, wherein the composition comprises from 50 to 200 mg ofthe oxygen-releasing agent.
 65. The composition of claim 44 or 45,wherein the composition comprises a dose of 5 mg to 100 mg of hydrogenperoxide equivalent.
 66. The composition of claim 44 or 45, wherein thecomposition comprises a dose of 20 mg to 40 mg of hydrogen peroxideequivalent.
 67. The composition of claim 44 or 45, wherein the weightratio of the enteric coating to the core of the particles is from 5% to50%.
 68. The composition of claim 44 or 45, wherein the particlescomprise a subcoat layer that encases the core and is disposed betweenthe core and the enteric coating.
 69. The composition of claim 68,wherein the subcoat layer comprises a release rate modifier.
 70. Thecomposition of claim 68, wherein the subcoat layer comprises HPMC. 71.The composition of claim 44 or 45, wherein the composition is formulatedfor oral administration and release of the oxygen-releasing agent fromthe particles in the human small intestine and/or colon.
 72. Thecomposition of claim 44 or 45, wherein the composition is formulated fororal administration and release of the oxygen-releasing agent from theparticles in the human ileum.
 73. The composition of claim 44 or 45,wherein the composition is formulated for oral administration andrelease of the oxygen-releasing agent from the particles in the humandistal ileum.
 74. The composition of claim 44 or 45, wherein thecomposition is formulated for oral administration and release of oxygenfrom the particles in the human small intestine and/or colon.
 75. Thecomposition of claim 44 or 45, wherein the composition is formulated fororal administration and release of oxygen from the particles in thehuman ileum.
 76. The composition of claim 44 or 45, wherein thecomposition is formulated for oral administration and release of oxygenfrom the particles in the human distal ileum.